Podcast
Questions and Answers
Which of the following substances is LEAST effectively demonstrated by the Periodic Acid-Schiff (PAS) stain?
Which of the following substances is LEAST effectively demonstrated by the Periodic Acid-Schiff (PAS) stain?
- Basement membranes
- Neutral mucopolysaccharides
- Glycogen
- Sulfated mucopolysaccharides (correct)
What is the primary purpose of using diastase in conjunction with the Periodic Acid-Schiff (PAS) stain?
What is the primary purpose of using diastase in conjunction with the Periodic Acid-Schiff (PAS) stain?
- To enhance the staining intensity of mucopolysaccharides
- To digest and remove glycogen, improving specificity (correct)
- To block non-specific binding of the Schiff reagent
- To oxidize carbohydrates more effectively
Which fixative is most desirable for Best Carmine staining to ensure optimal results?
Which fixative is most desirable for Best Carmine staining to ensure optimal results?
- Bouin's solution (correct)
- 10% neutral buffered formalin
- Carnoy's fixative
- Zenker's solution
What is the primary component stained by Mayer Mucicarmine?
What is the primary component stained by Mayer Mucicarmine?
For which of the following stains is maintaining the correct pH MOST critical for accurate results?
For which of the following stains is maintaining the correct pH MOST critical for accurate results?
If a lab is attempting to differentiate between hyaluronic acid and other acidic mucopolysaccharides, which technique would be MOST appropriate?
If a lab is attempting to differentiate between hyaluronic acid and other acidic mucopolysaccharides, which technique would be MOST appropriate?
You observe a staining result where acidic mucosubstances are blue and neutral polysaccharides are magenta. Which staining technique was MOST likely used?
You observe a staining result where acidic mucosubstances are blue and neutral polysaccharides are magenta. Which staining technique was MOST likely used?
Which of the following stains requires the use of polarized light microscopy for definitive identification of the target substance?
Which of the following stains requires the use of polarized light microscopy for definitive identification of the target substance?
Amyloid typically exhibits what characteristic appearance when stained with Congo Red and viewed under polarized light?
Amyloid typically exhibits what characteristic appearance when stained with Congo Red and viewed under polarized light?
Which of the following stains is used for visualizing amyloid fibrils using fluorescence microscopy?
Which of the following stains is used for visualizing amyloid fibrils using fluorescence microscopy?
1% acetic acid is used with what stain?
1% acetic acid is used with what stain?
Which of the following is a complex carbohydrate composed of long chains of monosaccharide units bonded together.
Which of the following is a complex carbohydrate composed of long chains of monosaccharide units bonded together.
If a tissue sample shows a change in color of a stain when it binds to certain tissue components, indicating the presence of specific molecules or structures, what is this phenomenon called?
If a tissue sample shows a change in color of a stain when it binds to certain tissue components, indicating the presence of specific molecules or structures, what is this phenomenon called?
In which type of tissue would you primarily find mucins involved in providing structural support and maintaining tissue hydration?
In which type of tissue would you primarily find mucins involved in providing structural support and maintaining tissue hydration?
What is the MOST likely reason for a Periodic Acid-Schiff (PAS) stain to show weak or absent staining of glycogen in a liver tissue control?
What is the MOST likely reason for a Periodic Acid-Schiff (PAS) stain to show weak or absent staining of glycogen in a liver tissue control?
In a PAS with diastase stain, if the tissue section shows persistent magenta staining even after diastase treatment, what is the MOST probable cause?
In a PAS with diastase stain, if the tissue section shows persistent magenta staining even after diastase treatment, what is the MOST probable cause?
During a Best Carmine stain, you observe a diffuse staining pattern with poor differentiation. What is the MOST likely cause?
During a Best Carmine stain, you observe a diffuse staining pattern with poor differentiation. What is the MOST likely cause?
In Mayer Mucicarmine staining, if the mucin does NOT stain properly, despite using a known positive control, what is the MOST likely source of error?
In Mayer Mucicarmine staining, if the mucin does NOT stain properly, despite using a known positive control, what is the MOST likely source of error?
You are performing Alcian Blue staining at pH 2.5, but the tissue section also contains sulfated mucopolysaccharides; what is the LIKELY staining result?
You are performing Alcian Blue staining at pH 2.5, but the tissue section also contains sulfated mucopolysaccharides; what is the LIKELY staining result?
If Alcian Blue staining at pH 1.0 yields NO staining, even with a known positive control, what is the MOST probable cause?
If Alcian Blue staining at pH 1.0 yields NO staining, even with a known positive control, what is the MOST probable cause?
In Müller-Mowry Colloidal Iron staining, what adjustment should be made if the background staining is excessively high?
In Müller-Mowry Colloidal Iron staining, what adjustment should be made if the background staining is excessively high?
Following Congo Red staining, a pathologist FAILS to observe apple-green birefringence in a tissue sample known to contain amyloid. What is the MOST likely reason?
Following Congo Red staining, a pathologist FAILS to observe apple-green birefringence in a tissue sample known to contain amyloid. What is the MOST likely reason?
Why is it important to use chemically clean glassware when performing special stains such as PAS, Best Carmine, or Alcian Blue?
Why is it important to use chemically clean glassware when performing special stains such as PAS, Best Carmine, or Alcian Blue?
After performing a Thioflavin T stain, the control tissue known to contain amyloid does NOT fluoresce. What is the MOST likely reason for this?
After performing a Thioflavin T stain, the control tissue known to contain amyloid does NOT fluoresce. What is the MOST likely reason for this?
A laboratory is switching from formalin to a different fixative for all tissue samples. How might this change affect the staining results for the special stains discussed?
A laboratory is switching from formalin to a different fixative for all tissue samples. How might this change affect the staining results for the special stains discussed?
In crystal violet staining, what cellular component facilitates the uptake and retention of the stain, resulting in the characteristic purple color?
In crystal violet staining, what cellular component facilitates the uptake and retention of the stain, resulting in the characteristic purple color?
A tissue sample stained with Alcian Blue at pH 2.5 shows minimal staining of acid mucopolysaccharides. Which of the following quality control measures should be implemented FIRST?
A tissue sample stained with Alcian Blue at pH 2.5 shows minimal staining of acid mucopolysaccharides. Which of the following quality control measures should be implemented FIRST?
Why is polarized light used in conjunction with Congo Red to identify amyloid deposits?
Why is polarized light used in conjunction with Congo Red to identify amyloid deposits?
What is the role of 10% neutral buffered formalin in the staining techniques?
What is the role of 10% neutral buffered formalin in the staining techniques?
What distinguishes epithelial mucins from connective tissue mucins in terms of their function?
What distinguishes epithelial mucins from connective tissue mucins in terms of their function?
What type of tissue serves as the BEST control material for Alcian Blue staining at pH 2.5?
What type of tissue serves as the BEST control material for Alcian Blue staining at pH 2.5?
What is the purpose of acetic acid, in the Thioflavin T stain?
What is the purpose of acetic acid, in the Thioflavin T stain?
What is the purpose of using control materials during special staining procedures?
What is the purpose of using control materials during special staining procedures?
A laboratory technologist notes that a newly prepared batch of Schiff reagent shows a pink hue even before use. What does it suggest the solution is?
A laboratory technologist notes that a newly prepared batch of Schiff reagent shows a pink hue even before use. What does it suggest the solution is?
Which of the following fiber types provides the MOST significant resistance to stretching forces in tendons and ligaments?
Which of the following fiber types provides the MOST significant resistance to stretching forces in tendons and ligaments?
What characteristic of cardiac muscle fibers allows for the rapid and coordinated contraction of the heart?
What characteristic of cardiac muscle fibers allows for the rapid and coordinated contraction of the heart?
What is the primary function of fibroblasts in connective tissue?
What is the primary function of fibroblasts in connective tissue?
Which cellular component is MOST directly responsible for the elasticity observed in lung tissue?
Which cellular component is MOST directly responsible for the elasticity observed in lung tissue?
Which of the following statements BEST describes the arrangement of reticular fibers within soft tissues?
Which of the following statements BEST describes the arrangement of reticular fibers within soft tissues?
Which characteristic BEST differentiates skeletal muscle fibers from smooth muscle fibers?
Which characteristic BEST differentiates skeletal muscle fibers from smooth muscle fibers?
Which of the following cell types is MOST active during an inflammatory response, engulfing pathogens and releasing cytokines?
Which of the following cell types is MOST active during an inflammatory response, engulfing pathogens and releasing cytokines?
In the Masson trichrome stain, what is the role of phosphomolybdic-phosphotungstic acid?
In the Masson trichrome stain, what is the role of phosphomolybdic-phosphotungstic acid?
If a Masson trichrome-stained tissue section shows weak collagen staining, what is the MOST appropriate initial corrective action?
If a Masson trichrome-stained tissue section shows weak collagen staining, what is the MOST appropriate initial corrective action?
Which component of Verhoeff's elastic stain acts as a mordant to enhance binding of the dye to elastic fibers?
Which component of Verhoeff's elastic stain acts as a mordant to enhance binding of the dye to elastic fibers?
During a Gomori Aldehyde Fuchsin stain, you observe that the elastic fibers are weakly stained. What is the MOST likely cause?
During a Gomori Aldehyde Fuchsin stain, you observe that the elastic fibers are weakly stained. What is the MOST likely cause?
What is the purpose of hydrochloric acid in the Orcein stain?
What is the purpose of hydrochloric acid in the Orcein stain?
A tissue section stained with Russell modification of the Movat pentachrome stain shows poor differentiation between muscle and collagen. Which reagent is MOST likely to be adjusted to correct this?
A tissue section stained with Russell modification of the Movat pentachrome stain shows poor differentiation between muscle and collagen. Which reagent is MOST likely to be adjusted to correct this?
In Gomori silver impregnation, what is the purpose of gold chloride?
In Gomori silver impregnation, what is the purpose of gold chloride?
Why is formic acid used in the Gordon and Sweets silver impregnation method?
Why is formic acid used in the Gordon and Sweets silver impregnation method?
What is the function of sodium borate in the Methenamine-Silver stain?
What is the function of sodium borate in the Methenamine-Silver stain?
What is the MOST likely reason Mallory PTAH staining shows weak or absent staining of muscle striations in a skeletal muscle tissue control?
What is the MOST likely reason Mallory PTAH staining shows weak or absent staining of muscle striations in a skeletal muscle tissue control?
For optimal results with Oil Red O stain, why is it critical to use frozen sections instead of paraffin-embedded tissue?
For optimal results with Oil Red O stain, why is it critical to use frozen sections instead of paraffin-embedded tissue?
What is the underlying principle behind Sudan Black B staining of lipids?
What is the underlying principle behind Sudan Black B staining of lipids?
When using osmium tetroxide as a stain, what chemical interaction leads to the blackening of lipids?
When using osmium tetroxide as a stain, what chemical interaction leads to the blackening of lipids?
Why is it important to perform a buffer wash of tissue fixed with glutaraldehyde before toluidine blue staining?
Why is it important to perform a buffer wash of tissue fixed with glutaraldehyde before toluidine blue staining?
What staining characteristic of mast cell granules is BEST highlighted by Toluidine blue?
What staining characteristic of mast cell granules is BEST highlighted by Toluidine blue?
Which fixative is BEST suited for methyl green-pyronin staining to ensure optimal differentiation of DNA and RNA?
Which fixative is BEST suited for methyl green-pyronin staining to ensure optimal differentiation of DNA and RNA?
In methyl green-pyronin staining, what is the mechanism by which methyl green selectively stains DNA?
In methyl green-pyronin staining, what is the mechanism by which methyl green selectively stains DNA?
What is the primary reason for using Masson trichrome stain in histological analysis?
What is the primary reason for using Masson trichrome stain in histological analysis?
What cellular feature makes mast cells readily identifiable with special stains like Toluidine blue?
What cellular feature makes mast cells readily identifiable with special stains like Toluidine blue?
Which cytoplasmic substance is SPECIFICALLY targeted and stained by pyronin?
Which cytoplasmic substance is SPECIFICALLY targeted and stained by pyronin?
In histological staining, what does the term 'impregnation' refer to?
In histological staining, what does the term 'impregnation' refer to?
Which of the following stains is frequently used to visualize Hepatitis B surface antigen and copper-associated proteins?
Which of the following stains is frequently used to visualize Hepatitis B surface antigen and copper-associated proteins?
Which of the listed techniques is BEST for demonstrating basement membranes?
Which of the listed techniques is BEST for demonstrating basement membranes?
Which of the following fixatives is MOST likely to negatively impact lipid staining if used prior to performing Oil Red O?
Which of the following fixatives is MOST likely to negatively impact lipid staining if used prior to performing Oil Red O?
When performing a Gomori Silver Impregnation stain, what step is MOST critical for achieving selective staining of reticular fibers and preventing non-specific silver deposition?
When performing a Gomori Silver Impregnation stain, what step is MOST critical for achieving selective staining of reticular fibers and preventing non-specific silver deposition?
For the van Gieson stain, what is the purpose of using picric acid?
For the van Gieson stain, what is the purpose of using picric acid?
In the Mallory PTAH stain, which tissue components are stained blue?
In the Mallory PTAH stain, which tissue components are stained blue?
Why should prolonged storage in formalin be avoided after fixation when planning to perform Oil Red O or Sudan Black B stains?
Why should prolonged storage in formalin be avoided after fixation when planning to perform Oil Red O or Sudan Black B stains?
Which of the following is a component of Verhoeff's elastic stain?
Which of the following is a component of Verhoeff's elastic stain?
Which of the following stains is used to demonstrate elastic fibers, mucopolysaccharides, and pancreatic beta cells?
Which of the following stains is used to demonstrate elastic fibers, mucopolysaccharides, and pancreatic beta cells?
Which of the following techniques uses a reducing agent to convert silver ions to visible silver particles?
Which of the following techniques uses a reducing agent to convert silver ions to visible silver particles?
Of the following options, which represents the BEST control material for the Gomori silver impregnation technique?
Of the following options, which represents the BEST control material for the Gomori silver impregnation technique?
Which of the following tissue components is LEAST likely to be directly produced or maintained by fibroblasts?
Which of the following tissue components is LEAST likely to be directly produced or maintained by fibroblasts?
What is the PRIMARY reason for using multiple dyes with differing affinities in trichrome staining methods?
What is the PRIMARY reason for using multiple dyes with differing affinities in trichrome staining methods?
In Verhoeff's elastic stain, what is the role of ferric chloride subsequent to staining with the hematoxylin-iodine solution?
In Verhoeff's elastic stain, what is the role of ferric chloride subsequent to staining with the hematoxylin-iodine solution?
A laboratory is experiencing inconsistent staining with Gomori Aldehyde Fuchsin. Which of the following is the MOST likely cause of this issue?
A laboratory is experiencing inconsistent staining with Gomori Aldehyde Fuchsin. Which of the following is the MOST likely cause of this issue?
What is the function of hydrochloric acid in the Orcein staining method?
What is the function of hydrochloric acid in the Orcein staining method?
In the Russell modification of Movat pentachrome stain, if collagen appears red instead of yellow, what adjustment to the staining procedure is MOST likely required?
In the Russell modification of Movat pentachrome stain, if collagen appears red instead of yellow, what adjustment to the staining procedure is MOST likely required?
Following Gomori silver impregnation, the reticular fibers in a tissue section appear faint and poorly defined. Which of the following is the MOST likely cause?
Following Gomori silver impregnation, the reticular fibers in a tissue section appear faint and poorly defined. Which of the following is the MOST likely cause?
What is the purpose of formic acid in the Gordon and Sweets silver impregnation method?
What is the purpose of formic acid in the Gordon and Sweets silver impregnation method?
In the Methenamine-Silver stain, what is the consequence of omitting sodium borate from the staining procedure?
In the Methenamine-Silver stain, what is the consequence of omitting sodium borate from the staining procedure?
What is the MOST critical consideration when using Mallory PTAH stain to ensure optimal staining of muscle striations?
What is the MOST critical consideration when using Mallory PTAH stain to ensure optimal staining of muscle striations?
What chemical property of lipids allows them to be stained by Oil Red O and Sudan Black B?
What chemical property of lipids allows them to be stained by Oil Red O and Sudan Black B?
In osmium tetroxide staining, what type of chemical bond is MOST directly involved in the blackening of lipids?
In osmium tetroxide staining, what type of chemical bond is MOST directly involved in the blackening of lipids?
When tissue fixed with glutaraldehyde is to be stained with toluidine blue, what is the purpose of performing a buffer wash prior to staining?
When tissue fixed with glutaraldehyde is to be stained with toluidine blue, what is the purpose of performing a buffer wash prior to staining?
Which characteristic of mast cell granules contributes MOST to their unique staining properties with Toluidine blue?
Which characteristic of mast cell granules contributes MOST to their unique staining properties with Toluidine blue?
What is the primary function of methyl green in the methyl green-pyronin staining method?
What is the primary function of methyl green in the methyl green-pyronin staining method?
What is the PRIMARY purpose of utilizing the Masson trichrome stain in histological analysis?
What is the PRIMARY purpose of utilizing the Masson trichrome stain in histological analysis?
What is the key feature of mast cells that allows them to be readily identified with special stains like Toluidine blue?
What is the key feature of mast cells that allows them to be readily identified with special stains like Toluidine blue?
In the context of histological staining, what does the term 'impregnation' MOST accurately describe?
In the context of histological staining, what does the term 'impregnation' MOST accurately describe?
Which staining method is BEST suited for demonstrating reticular fibers in tissues such as the spleen or lymph nodes?
Which staining method is BEST suited for demonstrating reticular fibers in tissues such as the spleen or lymph nodes?
Why is fresh frozen tissue generally required for Oil Red O and Sudan Black B staining?
Why is fresh frozen tissue generally required for Oil Red O and Sudan Black B staining?
Which cellular component is MOST directly associated with the function of Nissl substance?
Which cellular component is MOST directly associated with the function of Nissl substance?
What is the PRIMARY function of myelin sheath that surrounds certain nerve fibers?
What is the PRIMARY function of myelin sheath that surrounds certain nerve fibers?
Along what structure are impulses conducted away from the nerve cell body towards other cells?
Along what structure are impulses conducted away from the nerve cell body towards other cells?
If a tissue sample is fixed in a non-formalin fixative, which of the following actions is generally recommended before performing Cresyl echt violet staining?
If a tissue sample is fixed in a non-formalin fixative, which of the following actions is generally recommended before performing Cresyl echt violet staining?
What component within neurons does Cresyl echt violet specifically target for visualization?
What component within neurons does Cresyl echt violet specifically target for visualization?
What would be the MOST likely visual result of Cresyl echt violet staining on brain tissue?
What would be the MOST likely visual result of Cresyl echt violet staining on brain tissue?
Which of the following could cause inconsistent staining results with the Cresyl echt violet method?
Which of the following could cause inconsistent staining results with the Cresyl echt violet method?
How does Cresyl echt violet interact with neuronal tissue to produce a visible stain?
How does Cresyl echt violet interact with neuronal tissue to produce a visible stain?
What tissue component is highlighted by the Bodian stain?
What tissue component is highlighted by the Bodian stain?
If a Bodian stain exhibits incomplete impregnation of the target structures, what adjustment is MOST appropriate?
If a Bodian stain exhibits incomplete impregnation of the target structures, what adjustment is MOST appropriate?
What is the expected staining outcome for nerve fibers when using Holmes silver nitrate stain?
What is the expected staining outcome for nerve fibers when using Holmes silver nitrate stain?
Which staining method relies on silver binding to neurofibrils to visualize nerve fibers?
Which staining method relies on silver binding to neurofibrils to visualize nerve fibers?
Which stain is MOST similar to the Bielschowsky stain in terms of the tissue components it highlights?
Which stain is MOST similar to the Bielschowsky stain in terms of the tissue components it highlights?
What is the expected fluorescence of amyloid deposits when stained with Thioflavin S and viewed under UV light?
What is the expected fluorescence of amyloid deposits when stained with Thioflavin S and viewed under UV light?
What is the mechanism of action of Thioflavin S in staining amyloid?
What is the mechanism of action of Thioflavin S in staining amyloid?
What type of microscopy is required to visualize the results of Thioflavin S staining?
What type of microscopy is required to visualize the results of Thioflavin S staining?
What tissue components are stained blue by PTAH?
What tissue components are stained blue by PTAH?
What aspect of tissue processing is MOST crucial for optimal PTAH staining results?
What aspect of tissue processing is MOST crucial for optimal PTAH staining results?
If a PTAH stain shows overstaining, what corrective action should be taken?
If a PTAH stain shows overstaining, what corrective action should be taken?
What does Holzer staining primarily demonstrate in tissue sections?
What does Holzer staining primarily demonstrate in tissue sections?
What is the role of phosphomolybdic acid in Holzer staining?
What is the role of phosphomolybdic acid in Holzer staining?
What color do glial fibers typically appear after Holzer staining?
What color do glial fibers typically appear after Holzer staining?
What is the PRIMARY purpose of the Cajal stain in histology?
What is the PRIMARY purpose of the Cajal stain in histology?
Following Cajal staining, what color do astrocytes typically appear against the background?
Following Cajal staining, what color do astrocytes typically appear against the background?
What reagent is used in the Cajal stain to impregnate astrocytes?
What reagent is used in the Cajal stain to impregnate astrocytes?
Which fixative is MOST suitable for the Weil staining method?
Which fixative is MOST suitable for the Weil staining method?
What is the role of ferric ammonium sulfate in the Weil stain?
What is the role of ferric ammonium sulfate in the Weil stain?
Which component acts as a differentiator in Weil staining, ensuring clear visualization of myelin sheaths?
Which component acts as a differentiator in Weil staining, ensuring clear visualization of myelin sheaths?
What tissue component is stained by Luxol fast blue?
What tissue component is stained by Luxol fast blue?
In Luxol fast blue staining, what is the purpose of cresyl echt violet?
In Luxol fast blue staining, what is the purpose of cresyl echt violet?
With Luxol fast blue staining, what color will myelin appear?
With Luxol fast blue staining, what color will myelin appear?
Which of the following bacterial shapes is correctly matched with an example?
Which of the following bacterial shapes is correctly matched with an example?
Which of the following is a characteristic that differentiates mycobacteria from other types of bacteria?
Which of the following is a characteristic that differentiates mycobacteria from other types of bacteria?
Which of the following techniques is MOST appropriate for the initial detection of Mycobacterium tuberculosis in a tissue sample?
Which of the following techniques is MOST appropriate for the initial detection of Mycobacterium tuberculosis in a tissue sample?
In acid-fast staining, what is the role of the acid-alcohol solution?
In acid-fast staining, what is the role of the acid-alcohol solution?
Which of the following best describes the expected staining result of a Gram stain on a sample of Staphylococcus aureus?
Which of the following best describes the expected staining result of a Gram stain on a sample of Staphylococcus aureus?
Why is methanol or heat fixation preferred over formalin fixation when performing a Gram stain?
Why is methanol or heat fixation preferred over formalin fixation when performing a Gram stain?
What is the primary purpose of using iodine as a mordant in the Gram staining procedure?
What is the primary purpose of using iodine as a mordant in the Gram staining procedure?
A Giemsa stain is performed on a bone marrow sample. Which of the following organisms would this stain be MOST useful in identifying?
A Giemsa stain is performed on a bone marrow sample. Which of the following organisms would this stain be MOST useful in identifying?
How does Giemsa stain differentiate cellular structures?
How does Giemsa stain differentiate cellular structures?
Which component of Helicobacter pylori is targeted by Alcian yellow in the Alcian yellow-toluidine blue staining method?
Which component of Helicobacter pylori is targeted by Alcian yellow in the Alcian yellow-toluidine blue staining method?
In a PAS stain, what chemical process occurs when periodic acid is applied to the tissue section?
In a PAS stain, what chemical process occurs when periodic acid is applied to the tissue section?
Which type of organism is BEST demonstrated using the periodic acid-Schiff (PAS) stain?
Which type of organism is BEST demonstrated using the periodic acid-Schiff (PAS) stain?
Following chromic acid oxidation in the Chromic Acid-Schiff (CAS) stain, what functional group is created in fungal cell walls that allows for subsequent staining?
Following chromic acid oxidation in the Chromic Acid-Schiff (CAS) stain, what functional group is created in fungal cell walls that allows for subsequent staining?
What is the role of aldehyde fuchsin in the Gridley fungus stain?
What is the role of aldehyde fuchsin in the Gridley fungus stain?
Which of the following reagents is used to tone the silver stain in the Grocott modification of Gomori methenamine-silver (GMS) method?
Which of the following reagents is used to tone the silver stain in the Grocott modification of Gomori methenamine-silver (GMS) method?
In the Grocott modification of Gomori methenamine-silver stain, what is the purpose of sodium bisulfite?
In the Grocott modification of Gomori methenamine-silver stain, what is the purpose of sodium bisulfite?
What component of Cryptococcus species does Mayer mucicarmine specifically stain?
What component of Cryptococcus species does Mayer mucicarmine specifically stain?
In the Warthin-Starry stain, what process occurs that makes spirochetes visible?
In the Warthin-Starry stain, what process occurs that makes spirochetes visible?
Hydroquinone is used in silver impregnation techniques such as the Warthin-Starry and Dieterle stains. What is its function?
Hydroquinone is used in silver impregnation techniques such as the Warthin-Starry and Dieterle stains. What is its function?
Which of the following reagents acts as a sensitizer in the Dieterle stain, enhancing silver impregnation of microorganisms?
Which of the following reagents acts as a sensitizer in the Dieterle stain, enhancing silver impregnation of microorganisms?
In the Steiner and Steiner stain, what is the role of silver nitrate?
In the Steiner and Steiner stain, what is the role of silver nitrate?
Which of the following fixatives is MOST desirable for preserving Mycobacterium tuberculosis in tissue samples prior to performing acid-fast staining?
Which of the following fixatives is MOST desirable for preserving Mycobacterium tuberculosis in tissue samples prior to performing acid-fast staining?
What is the MOST critical step to ensure accurate and reliable results when performing the Auramine-rhodamine stain for mycobacteria?
What is the MOST critical step to ensure accurate and reliable results when performing the Auramine-rhodamine stain for mycobacteria?
If a Gram-stained tissue section shows excessive decolorization, causing Gram-positive bacteria to appear pink instead of purple, which of the following corrective actions is MOST appropriate?
If a Gram-stained tissue section shows excessive decolorization, causing Gram-positive bacteria to appear pink instead of purple, which of the following corrective actions is MOST appropriate?
During a Giemsa staining procedure, a technologist observes that the cellular details are poorly defined and the overall staining intensity is weak. What is the MOST likely cause?
During a Giemsa staining procedure, a technologist observes that the cellular details are poorly defined and the overall staining intensity is weak. What is the MOST likely cause?
A tissue sample stained with Alcian yellow-toluidine blue shows a very pale yellow staining of Helicobacter pylori, making it difficult to visualize. What is the MOST probable cause?
A tissue sample stained with Alcian yellow-toluidine blue shows a very pale yellow staining of Helicobacter pylori, making it difficult to visualize. What is the MOST probable cause?
After performing a PAS stain, the control tissue shows very weak magenta staining. What is the MOST likely cause?
After performing a PAS stain, the control tissue shows very weak magenta staining. What is the MOST likely cause?
Fungal elements are not visible in a tissue known to contain Histoplasma species. Which step in the Chromic Acid-Schiff (CAS) staining procedure is the MOST likely source of error?
Fungal elements are not visible in a tissue known to contain Histoplasma species. Which step in the Chromic Acid-Schiff (CAS) staining procedure is the MOST likely source of error?
A Gridley fungus stain demonstrates weak staining of fungal elements. What adjustment to the staining procedure is MOST likely to correct this issue?
A Gridley fungus stain demonstrates weak staining of fungal elements. What adjustment to the staining procedure is MOST likely to correct this issue?
After performing the Grocott methenamine-silver (GMS) stain, the tissue section appears overstained, with a dark, non-specific staining pattern. What is the MOST likely cause of this issue?
After performing the Grocott methenamine-silver (GMS) stain, the tissue section appears overstained, with a dark, non-specific staining pattern. What is the MOST likely cause of this issue?
In Mayer mucicarmine staining, if the capsules of Cryptococcus species are not clearly stained red, despite appropriate timing and technique, what is the MOST probable cause?
In Mayer mucicarmine staining, if the capsules of Cryptococcus species are not clearly stained red, despite appropriate timing and technique, what is the MOST probable cause?
If a Warthin-Starry stained tissue section shows excessive background staining, obscuring the spirochetes, what corrective action should be taken FIRST?
If a Warthin-Starry stained tissue section shows excessive background staining, obscuring the spirochetes, what corrective action should be taken FIRST?
Which of the following modifications to the Dieterle stain is most likely to improve the visualization of Legionella species in lung tissue?
Which of the following modifications to the Dieterle stain is most likely to improve the visualization of Legionella species in lung tissue?
During a Steiner and Steiner staining procedure, the control tissue known to contain spirochetes shows very weak or absent staining. What is the MOST likely explanation?
During a Steiner and Steiner staining procedure, the control tissue known to contain spirochetes shows very weak or absent staining. What is the MOST likely explanation?
Which staining technique uses a combination of fluorescent dyes to detect mycobacteria?
Which staining technique uses a combination of fluorescent dyes to detect mycobacteria?
What is the purpose of potassium permanganate in the Auramine-rhodamine staining method?
What is the purpose of potassium permanganate in the Auramine-rhodamine staining method?
If a Ziehl-Neelsen stain for acid-fast bacteria is too pale, and the technologist suspects the carbol fuchsin stain is not penetrating the cell walls effectively, what modification to the procedure might improve the staining?
If a Ziehl-Neelsen stain for acid-fast bacteria is too pale, and the technologist suspects the carbol fuchsin stain is not penetrating the cell walls effectively, what modification to the procedure might improve the staining?
When performing a Gram stain, which step is MOST critical in preventing a false-negative result (i.e., Gram-positive bacteria appearing Gram-negative)?
When performing a Gram stain, which step is MOST critical in preventing a false-negative result (i.e., Gram-positive bacteria appearing Gram-negative)?
A pathologist requests a Giemsa stain on a peripheral blood smear to evaluate for malaria parasites. If the parasites are not clearly visible, despite the use of a known positive control, what is the MOST probable cause?
A pathologist requests a Giemsa stain on a peripheral blood smear to evaluate for malaria parasites. If the parasites are not clearly visible, despite the use of a known positive control, what is the MOST probable cause?
A biopsy from the stomach is stained with Alcian Yellow-Toluidine Blue to check for Helicobacter pylori. The background is intensely stained blue, obscuring any potential bacteria. What step can MOST likely be modified to improve visualization?
A biopsy from the stomach is stained with Alcian Yellow-Toluidine Blue to check for Helicobacter pylori. The background is intensely stained blue, obscuring any potential bacteria. What step can MOST likely be modified to improve visualization?
A surgical specimen is stained with PAS to identify a fungal infection, but only a weak, diffuse staining pattern is observed. To improve the results, which step in the staining procedure should be optimized FIRST?
A surgical specimen is stained with PAS to identify a fungal infection, but only a weak, diffuse staining pattern is observed. To improve the results, which step in the staining procedure should be optimized FIRST?
A Grocott methenamine-silver (GMS) stain on lung tissue shows adequate staining of Pneumocystis jirovecii cysts, but there is also significant black precipitate deposited nonspecifically throughout the tissue. What is the MOST likely cause of this artifact?
A Grocott methenamine-silver (GMS) stain on lung tissue shows adequate staining of Pneumocystis jirovecii cysts, but there is also significant black precipitate deposited nonspecifically throughout the tissue. What is the MOST likely cause of this artifact?
A tissue section stained with Warthin-Starry demonstrates poor impregnation of spirochetes, and they appear faint and difficult to visualize. Which parameter should be adjusted to improve?
A tissue section stained with Warthin-Starry demonstrates poor impregnation of spirochetes, and they appear faint and difficult to visualize. Which parameter should be adjusted to improve?
After performing a Dieterle stain, a lab technician observes a high level of background staining, making it difficult to identify the target microorganisms. Which of the following actions would BEST address this issue?
After performing a Dieterle stain, a lab technician observes a high level of background staining, making it difficult to identify the target microorganisms. Which of the following actions would BEST address this issue?
A tissue sample suspected of containing Mycobacterium tuberculosis is fixed in Bouin's solution. What is the MOST appropriate next step before performing an acid-fast stain?
A tissue sample suspected of containing Mycobacterium tuberculosis is fixed in Bouin's solution. What is the MOST appropriate next step before performing an acid-fast stain?
A technologist performs an Auramine-rhodamine stain, but the mycobacteria are difficult to visualize against a bright background. What reagent adjustment would BEST address this issue?
A technologist performs an Auramine-rhodamine stain, but the mycobacteria are difficult to visualize against a bright background. What reagent adjustment would BEST address this issue?
During a Gram staining procedure, after applying crystal violet and iodine, the tissue section is rinsed with alcohol for an extended period. What is the MOST likely consequence of this over-decolorization?
During a Gram staining procedure, after applying crystal violet and iodine, the tissue section is rinsed with alcohol for an extended period. What is the MOST likely consequence of this over-decolorization?
A Giemsa stain on a bone marrow aspirate shows poor differentiation of cellular details. Which of the following is the MOST likely cause?
A Giemsa stain on a bone marrow aspirate shows poor differentiation of cellular details. Which of the following is the MOST likely cause?
A gastric biopsy is stained with Alcian yellow-toluidine blue but the Helicobacter pylori are difficult to visualize due to weak staining. Which of the following steps is MOST likely to improve the staining?
A gastric biopsy is stained with Alcian yellow-toluidine blue but the Helicobacter pylori are difficult to visualize due to weak staining. Which of the following steps is MOST likely to improve the staining?
A PAS stain on liver tissue shows weak staining of glycogen. Knowing that glycogen should be present, how should the staining procedure be modified to improve the result?
A PAS stain on liver tissue shows weak staining of glycogen. Knowing that glycogen should be present, how should the staining procedure be modified to improve the result?
In a Chromic Acid-Schiff (CAS) stain, if fungal elements are not visible in a tissue known to contain Histoplasma species, despite using a known positive control, the MOST probable cause is:
In a Chromic Acid-Schiff (CAS) stain, if fungal elements are not visible in a tissue known to contain Histoplasma species, despite using a known positive control, the MOST probable cause is:
After performing a Grocott methenamine-silver (GMS) stain, the tissue section is overstained with a dark, non-specific staining pattern. Which of the following steps would BEST correct this issue?
After performing a Grocott methenamine-silver (GMS) stain, the tissue section is overstained with a dark, non-specific staining pattern. Which of the following steps would BEST correct this issue?
A Warthin-Starry stain demonstrates significant background staining, obscuring any spirochetes. What corrective action should be taken FIRST?
A Warthin-Starry stain demonstrates significant background staining, obscuring any spirochetes. What corrective action should be taken FIRST?
If a Dieterle-stained tissue section shows very weak or absent staining of spirochetes in the control tissue known to contain them, which of the following is the MOST likely explanation?
If a Dieterle-stained tissue section shows very weak or absent staining of spirochetes in the control tissue known to contain them, which of the following is the MOST likely explanation?
Which of the following is an example of an endogenous pigment?
Which of the following is an example of an endogenous pigment?
Which pigment is classified as both exogenous and specifically linked to inhalation of particulate matter?
Which pigment is classified as both exogenous and specifically linked to inhalation of particulate matter?
Which of the following pigments is derived from the breakdown of erythrocytes?
Which of the following pigments is derived from the breakdown of erythrocytes?
What distinguishes an argentaffin reaction from an argyrophil reaction?
What distinguishes an argentaffin reaction from an argyrophil reaction?
In which type of tissue or cells would you expect to find argentaffin granules that can be demonstrated without an external reducing agent?
In which type of tissue or cells would you expect to find argentaffin granules that can be demonstrated without an external reducing agent?
Which of the following techniques relies on the use of an external reducing agent to visualize the target substance?
Which of the following techniques relies on the use of an external reducing agent to visualize the target substance?
Which substance is demonstrated by the Prussian blue staining technique?
Which substance is demonstrated by the Prussian blue staining technique?
If a lab technician needs to confirm the presence of hemosiderin in a tissue sample, which of the following stains should be used?
If a lab technician needs to confirm the presence of hemosiderin in a tissue sample, which of the following stains should be used?
What type of substance is demonstrated by the Schmorl ferric-ferricyanide reduction test?
What type of substance is demonstrated by the Schmorl ferric-ferricyanide reduction test?
A pathologist suspects a tumor has the capability of reducing silver ions without needing an external reducing agent. Which stain would be most appropriate for this assessment?
A pathologist suspects a tumor has the capability of reducing silver ions without needing an external reducing agent. Which stain would be most appropriate for this assessment?
Which of the following staining techniques is used to identify argyrophil cells in a tissue section?
Which of the following staining techniques is used to identify argyrophil cells in a tissue section?
What is the target substance demonstrated by the Gomori methenamine-silver (GMS) stain?
What is the target substance demonstrated by the Gomori methenamine-silver (GMS) stain?
If a pathologist suspects a patient has a buildup of bilirubin in their liver tissue, which stain would be MOST appropriate to confirm this?
If a pathologist suspects a patient has a buildup of bilirubin in their liver tissue, which stain would be MOST appropriate to confirm this?
Which of the following techniques is used to demonstrate calcium deposits within a tissue sample?
Which of the following techniques is used to demonstrate calcium deposits within a tissue sample?
What is the purpose of using Alizarin red S in histopathology?
What is the purpose of using Alizarin red S in histopathology?
Which staining method is used to identify and visualize copper deposits in liver tissue?
Which staining method is used to identify and visualize copper deposits in liver tissue?
Why are alcohol-based fixatives or neutral buffered formalin considered the MOST desirable fixatives for the Prussian blue reaction?
Why are alcohol-based fixatives or neutral buffered formalin considered the MOST desirable fixatives for the Prussian blue reaction?
What chemical reaction occurs during the Prussian blue staining procedure that leads to the formation of the characteristic blue color?
What chemical reaction occurs during the Prussian blue staining procedure that leads to the formation of the characteristic blue color?
What is the MOST likely consequence of inadequate washing after fixation in the Schmorl ferric-ferricyanide reduction test?
What is the MOST likely consequence of inadequate washing after fixation in the Schmorl ferric-ferricyanide reduction test?
What step is critical in the Fontana-Masson stain to prevent non-specific staining caused by overexposure to silver nitrate?
What step is critical in the Fontana-Masson stain to prevent non-specific staining caused by overexposure to silver nitrate?
A tissue sample stained with the Grimelius method exhibits excessive background staining. What is the MOST likely reason for this?
A tissue sample stained with the Grimelius method exhibits excessive background staining. What is the MOST likely reason for this?
What is the purpose of using chromic acid in the Gomori methenamine-silver (GMS) stain?
What is the purpose of using chromic acid in the Gomori methenamine-silver (GMS) stain?
During a Hall (Fouchet) stain, a lab technician observes a loss of bilirubin staining. What could have caused this?
During a Hall (Fouchet) stain, a lab technician observes a loss of bilirubin staining. What could have caused this?
What might cause the lack of staining in a von Kossa stain if the appropriate control tissue is also negative?
What might cause the lack of staining in a von Kossa stain if the appropriate control tissue is also negative?
When performing an Alizarin red S stain, what step is critical to ensure optimal staining of calcium deposits?
When performing an Alizarin red S stain, what step is critical to ensure optimal staining of calcium deposits?
A laboratory technician is performing a Rhodanine stain and observes weak staining of copper deposits. What is the MOST probable cause?
A laboratory technician is performing a Rhodanine stain and observes weak staining of copper deposits. What is the MOST probable cause?
What is the primary difference in the chemical reactions between the Prussian blue and Turnbull blue reactions?
What is the primary difference in the chemical reactions between the Prussian blue and Turnbull blue reactions?
During iron staining, why is it important to differentiate between ferric and ferrous iron?
During iron staining, why is it important to differentiate between ferric and ferrous iron?
In what part of the eye is melanin typically found, contributing to its color and protecting against UV radiation?
In what part of the eye is melanin typically found, contributing to its color and protecting against UV radiation?
Besides melanin, what other pigment contributes to the color of hair?
Besides melanin, what other pigment contributes to the color of hair?
Which method is specifically used to demonstrate calcium oxalate crystals in tissue sections?
Which method is specifically used to demonstrate calcium oxalate crystals in tissue sections?
If a pathologist suspects copper deposits that are not well-demonstrated by Rhodanine staining, which alternative method could be employed?
If a pathologist suspects copper deposits that are not well-demonstrated by Rhodanine staining, which alternative method could be employed?
Which method is used to specifically identify urate crystals in a tissue sample, often useful in diagnosing gout?
Which method is used to specifically identify urate crystals in a tissue sample, often useful in diagnosing gout?
Which of the following staining techniques is BEST for demonstrating amyloid deposits in tissues, especially when viewed under polarized light?
Which of the following staining techniques is BEST for demonstrating amyloid deposits in tissues, especially when viewed under polarized light?
Why is it important to hydrate tissue sections through graded alcohols to water as a pre-treatment step for bleaching melanin pigment?
Why is it important to hydrate tissue sections through graded alcohols to water as a pre-treatment step for bleaching melanin pigment?
Following bleaching of melanin pigment with potassium permanganate, what reagent is used to remove the brown precipitate formed?
Following bleaching of melanin pigment with potassium permanganate, what reagent is used to remove the brown precipitate formed?
How can anthracotic pigment be differentiated from formalin pigment in a tissue section?
How can anthracotic pigment be differentiated from formalin pigment in a tissue section?
A pathologist observes a dark brown pigment in a tissue sample. The pigment bleaches with hydrogen peroxide. What is the MOST likely identity of the pigment?
A pathologist observes a dark brown pigment in a tissue sample. The pigment bleaches with hydrogen peroxide. What is the MOST likely identity of the pigment?
Which pigment is both endogenous and nonhematogenous?
Which pigment is both endogenous and nonhematogenous?
What key characteristic differentiates the argentaffin reaction from the argyrophil reaction in staining?
What key characteristic differentiates the argentaffin reaction from the argyrophil reaction in staining?
Which staining technique is utilized to identify ferrous iron (Fe²⁺) in tissue samples?
Which staining technique is utilized to identify ferrous iron (Fe²⁺) in tissue samples?
For the Prussian blue staining technique, which fixative provides the most desirable results in preserving ferric iron?
For the Prussian blue staining technique, which fixative provides the most desirable results in preserving ferric iron?
In the Schmorl ferric-ferricyanide reduction test, what is the MOST critical step to prevent nonspecific staining?
In the Schmorl ferric-ferricyanide reduction test, what is the MOST critical step to prevent nonspecific staining?
What is a potential source of error in the Fontana-Masson stain that can lead to non-specific staining?
What is a potential source of error in the Fontana-Masson stain that can lead to non-specific staining?
A pathologist needs to demonstrate argyrophil granules. From the following list which is the MOST appropriate staining method to employ?
A pathologist needs to demonstrate argyrophil granules. From the following list which is the MOST appropriate staining method to employ?
Which of the following is an appropriate control material for the Gomori methenamine-silver (GMS) stain?
Which of the following is an appropriate control material for the Gomori methenamine-silver (GMS) stain?
What is the MOST appropriate action to take if a tissue sample is fixed with a non-formalin fixative and needs to be stained with the Prussian blue technique?
What is the MOST appropriate action to take if a tissue sample is fixed with a non-formalin fixative and needs to be stained with the Prussian blue technique?
A lung biopsy from a patient with a history of smoking shows black deposits. Which of the following tests would BEST differentiate anthracotic pigment from melanin?
A lung biopsy from a patient with a history of smoking shows black deposits. Which of the following tests would BEST differentiate anthracotic pigment from melanin?
What is the function of a chromogen in immunohistochemistry?
What is the function of a chromogen in immunohistochemistry?
Which characteristic distinguishes monoclonal antibodies from polyclonal antisera?
Which characteristic distinguishes monoclonal antibodies from polyclonal antisera?
In the context of immunohistochemistry, what is a key advantage of the indirect method compared to the direct method?
In the context of immunohistochemistry, what is a key advantage of the indirect method compared to the direct method?
What advantage does a polymeric detection system offer over other immunohistochemical techniques like ABC?
What advantage does a polymeric detection system offer over other immunohistochemical techniques like ABC?
Which parameter is MOST critical when preparing a specimen for immunofluorescence to ensure optimal antibody penetration?
Which parameter is MOST critical when preparing a specimen for immunofluorescence to ensure optimal antibody penetration?
Why is it important to avoid formaldehyde-based fixatives when preparing specimens for immunofluorescence?
Why is it important to avoid formaldehyde-based fixatives when preparing specimens for immunofluorescence?
How does protein blocking contribute to accurate immunohistochemical staining?
How does protein blocking contribute to accurate immunohistochemical staining?
Which immunoglobulin heavy chain is predominantly associated with allergic reactions?
Which immunoglobulin heavy chain is predominantly associated with allergic reactions?
What is the primary purpose of using negative tissue controls in immunohistochemistry?
What is the primary purpose of using negative tissue controls in immunohistochemistry?
In immunohistochemical staining, how would you address the issue of high background staining?
In immunohistochemical staining, how would you address the issue of high background staining?
Why is preparing positive controls within the laboratory preferred over purchasing them?
Why is preparing positive controls within the laboratory preferred over purchasing them?
During Heat-Induced Epitope Retrieval (HIER), what is the primary function of the buffer solution?
During Heat-Induced Epitope Retrieval (HIER), what is the primary function of the buffer solution?
When performing Heat-Induced Epitope Retrieval (HIER), why is it important to select an appropriate heating method such as microwave, pressure cooker, or water bath?
When performing Heat-Induced Epitope Retrieval (HIER), why is it important to select an appropriate heating method such as microwave, pressure cooker, or water bath?
Which factor is MOST critical when choosing a solution for enzyme-induced epitope retrieval (EIER)?
Which factor is MOST critical when choosing a solution for enzyme-induced epitope retrieval (EIER)?
Why is it important to consider the solubility properties of chromogens used in immunohistochemistry?
Why is it important to consider the solubility properties of chromogens used in immunohistochemistry?
What is the purpose of using chemicals like imidazole or nickel chloride to intensify the 3,3'-diaminobenzidine (DAB) reaction?
What is the purpose of using chemicals like imidazole or nickel chloride to intensify the 3,3'-diaminobenzidine (DAB) reaction?
Which factor MOST significantly contributes to the increased sensitivity observed in indirect immunohistochemical staining methods compared to direct methods?
Which factor MOST significantly contributes to the increased sensitivity observed in indirect immunohistochemical staining methods compared to direct methods?
A laboratory is experiencing inconsistent staining results with a particular antibody. If troubleshooting reveals no issues with the antibody itself or the tissue processing, which aspect of the immunohistochemical protocol should be examined NEXT?
A laboratory is experiencing inconsistent staining results with a particular antibody. If troubleshooting reveals no issues with the antibody itself or the tissue processing, which aspect of the immunohistochemical protocol should be examined NEXT?
In a diagnostic setting, when would monoclonal antibodies be preferred over polyclonal antisera for immunohistochemical staining?
In a diagnostic setting, when would monoclonal antibodies be preferred over polyclonal antisera for immunohistochemical staining?
What is the MOST likely reason for a technologist to choose an alcohol-based fixative over formalin when preparing a tissue sample for immunohistochemistry?
What is the MOST likely reason for a technologist to choose an alcohol-based fixative over formalin when preparing a tissue sample for immunohistochemistry?
Consider a scenario where the target antigen is present in very low concentrations within a tissue sample. Which detection method is MOST advisable?
Consider a scenario where the target antigen is present in very low concentrations within a tissue sample. Which detection method is MOST advisable?
What is the function of antibodies in the body?
What is the function of antibodies in the body?
What is the specific region of an antigen molecule to which an antibody binds?
What is the specific region of an antigen molecule to which an antibody binds?
What is the function of Multilink Secondary Antiserum?
What is the function of Multilink Secondary Antiserum?
Which of the following is an advantage of monoclonal antibodies over polyclonal antisera?
Which of the following is an advantage of monoclonal antibodies over polyclonal antisera?
Which type of detection method is commonly used in research and diagnostic labs for detecting a wide range of antigens, but requires a longer and more complex procedure?
Which type of detection method is commonly used in research and diagnostic labs for detecting a wide range of antigens, but requires a longer and more complex procedure?
Which of the following fluorochromes is commonly used in immunofluorescence and flow cytometry and emits a green fluorescence?
Which of the following fluorochromes is commonly used in immunofluorescence and flow cytometry and emits a green fluorescence?
When using immunoalkaline phosphatase techniques, which of the following reagents will show a magenta precipitate?
When using immunoalkaline phosphatase techniques, which of the following reagents will show a magenta precipitate?
Which enzyme system is classified under enzyme immunohistochemistry?
Which enzyme system is classified under enzyme immunohistochemistry?
Which of the following fixatives should be avoided when preparing a specimen for immunofluorescence?
Which of the following fixatives should be avoided when preparing a specimen for immunofluorescence?
What can you do to overcome the problem of cross-linking when using formaldehyde?
What can you do to overcome the problem of cross-linking when using formaldehyde?
Which non-formalin fixative poses an issue of causing autofluorescence?
Which non-formalin fixative poses an issue of causing autofluorescence?
How can the activity of endogenous peroxidase enzymes that cause non-specific staining be inhibited?
How can the activity of endogenous peroxidase enzymes that cause non-specific staining be inhibited?
What role does IgG play in the body?
What role does IgG play in the body?
What does it mean if staining is observed in the negative control tissue?
What does it mean if staining is observed in the negative control tissue?
Which of the following statements describe a corrective action for weak or no staining?
Which of the following statements describe a corrective action for weak or no staining?
Why is it important for lab-prepared positive controls to be tested with the test samples?
Why is it important for lab-prepared positive controls to be tested with the test samples?
Which of the following is a solution used for heat-induced epitope retrieval?
Which of the following is a solution used for heat-induced epitope retrieval?
When using a microwave during HIER, what's an important consideration?
When using a microwave during HIER, what's an important consideration?
Select the solution used for EIER that consists of a broad-sprectrum proteolytic enzyme
Select the solution used for EIER that consists of a broad-sprectrum proteolytic enzyme
Which of the following chemicals are used with Orange G?
Which of the following chemicals are used with Orange G?
If your DAB reaction needs to be darker, which of the following chemicals is MOST effective?
If your DAB reaction needs to be darker, which of the following chemicals is MOST effective?
Which of the following best describes the function of a chromogen in immunohistochemistry?
Which of the following best describes the function of a chromogen in immunohistochemistry?
What is the key difference between polyclonal and monoclonal antibodies regarding epitope recognition?
What is the key difference between polyclonal and monoclonal antibodies regarding epitope recognition?
In enzyme immunohistochemistry, which component directly results in the formation of a visible reaction product?
In enzyme immunohistochemistry, which component directly results in the formation of a visible reaction product?
Which of the following immunohistochemical techniques is LEAST likely to be affected by endogenous biotin?
Which of the following immunohistochemical techniques is LEAST likely to be affected by endogenous biotin?
Compared to direct immunohistochemistry, what is the primary advantage of using an indirect method?
Compared to direct immunohistochemistry, what is the primary advantage of using an indirect method?
If a researcher needs to visualize multiple antigens simultaneously in a tissue section, which technique would be the MOST appropriate?
If a researcher needs to visualize multiple antigens simultaneously in a tissue section, which technique would be the MOST appropriate?
For immunofluorescence staining, why is it recommended to avoid prolonged fixation in formaldehyde?
For immunofluorescence staining, why is it recommended to avoid prolonged fixation in formaldehyde?
In immunohistochemistry, what is the PRIMARY purpose of using a blocking solution?
In immunohistochemistry, what is the PRIMARY purpose of using a blocking solution?
What is the role of heat in heat-induced epitope retrieval (HIER)?
What is the role of heat in heat-induced epitope retrieval (HIER)?
What is the MOST likely consequence of omitting the permeabilization step when preparing a specimen for immunofluorescence?
What is the MOST likely consequence of omitting the permeabilization step when preparing a specimen for immunofluorescence?
A tissue section exhibits strong staining at the edges but minimal staining in the center. What is the MOST probable cause of this issue?
A tissue section exhibits strong staining at the edges but minimal staining in the center. What is the MOST probable cause of this issue?
Why are laboratory-prepared positive controls generally favored over commercially purchased ones for immunohistochemistry assays?
Why are laboratory-prepared positive controls generally favored over commercially purchased ones for immunohistochemistry assays?
What distinguishes alkaline phosphatase from horseradish peroxidase in enzyme immunohistochemistry?
What distinguishes alkaline phosphatase from horseradish peroxidase in enzyme immunohistochemistry?
Which of the following buffers is MOST commonly used for heat-induced epitope retrieval (HIER) at a high pH?
Which of the following buffers is MOST commonly used for heat-induced epitope retrieval (HIER) at a high pH?
If a DAB-stained slide appears weak, which of the following chemicals can be used to intensify the reaction?
If a DAB-stained slide appears weak, which of the following chemicals can be used to intensify the reaction?
Which of the following BEST describes the role of enzymes in biological reactions?
Which of the following BEST describes the role of enzymes in biological reactions?
Which of the following processes BEST exemplifies oxidation in a biological system?
Which of the following processes BEST exemplifies oxidation in a biological system?
In the context of enzyme-substrate interactions, what is the MOST accurate definition of a substrate?
In the context of enzyme-substrate interactions, what is the MOST accurate definition of a substrate?
Why is pH a critical factor when demonstrating enzyme activity through histochemistry?
Why is pH a critical factor when demonstrating enzyme activity through histochemistry?
Which of the following BEST describes the function of hydrolases?
Which of the following BEST describes the function of hydrolases?
In enzyme histochemistry, what is meant by 'simultaneous coupling'?
In enzyme histochemistry, what is meant by 'simultaneous coupling'?
What is the primary role of esterases in biochemical reactions?
What is the primary role of esterases in biochemical reactions?
Which of the following BEST describes the function of phosphatases?
Which of the following BEST describes the function of phosphatases?
What is the function of phosphorylases in biochemical pathways?
What is the function of phosphorylases in biochemical pathways?
Which connective tissue layer directly surrounds individual muscle fibers?
Which connective tissue layer directly surrounds individual muscle fibers?
What is the primary function of the epimysium in muscle tissue?
What is the primary function of the epimysium in muscle tissue?
What is the primary role of the perimysium within muscle tissue?
What is the primary role of the perimysium within muscle tissue?
Which of the following describes SKELETAL muscle fibers?
Which of the following describes SKELETAL muscle fibers?
Which structural feature is characteristic of CARDIAC muscle tissue?
Which structural feature is characteristic of CARDIAC muscle tissue?
Which feature distinguishes smooth muscle from skeletal and cardiac muscle?
Which feature distinguishes smooth muscle from skeletal and cardiac muscle?
What metabolic process primarily fuels type I muscle fibers?
What metabolic process primarily fuels type I muscle fibers?
Which characteristic is MOST indicative of type II muscle fibers?
Which characteristic is MOST indicative of type II muscle fibers?
Which muscle fiber type is BEST suited for endurance activities?
Which muscle fiber type is BEST suited for endurance activities?
What is a key characteristic that distinguishes enzymes from other catalysts?
What is a key characteristic that distinguishes enzymes from other catalysts?
How do enzymes enhance the rate of biochemical reactions?
How do enzymes enhance the rate of biochemical reactions?
Which of the following is a way in which enzyme activity can be regulated?
Which of the following is a way in which enzyme activity can be regulated?
What is the effect of increasing temperature on enzyme activity, assuming other factors are constant?
What is the effect of increasing temperature on enzyme activity, assuming other factors are constant?
How does increasing substrate concentration typically affect an enzyme-catalyzed reaction rate?
How does increasing substrate concentration typically affect an enzyme-catalyzed reaction rate?
What is the role of cofactors and coenzymes in enzyme function?
What is the role of cofactors and coenzymes in enzyme function?
What is the MOST common artifact seen in unfixed frozen tissue sections?
What is the MOST common artifact seen in unfixed frozen tissue sections?
Why is a cryoprotective solution used when storing tissue for enzyme studies?
Why is a cryoprotective solution used when storing tissue for enzyme studies?
Why is snap-freezing in liquid nitrogen the preferred method for freezing muscle tissue for enzyme studies?
Why is snap-freezing in liquid nitrogen the preferred method for freezing muscle tissue for enzyme studies?
What type of reaction underlies the α-Naphthyl Acetate Esterase technique?
What type of reaction underlies the α-Naphthyl Acetate Esterase technique?
What is the role of diazonium salt in the α-Naphthyl Acetate Esterase reaction?
What is the role of diazonium salt in the α-Naphthyl Acetate Esterase reaction?
What is the basis for visualizing ATPase activity in histochemical techniques?
What is the basis for visualizing ATPase activity in histochemical techniques?
Which of the following statements BEST describes the purpose of using a diazonium salt in the naphthol AS-D chloroacetate esterase stain?
Which of the following statements BEST describes the purpose of using a diazonium salt in the naphthol AS-D chloroacetate esterase stain?
In acid phosphatase reactions, what is the role of the substrate, such as fast red TR?
In acid phosphatase reactions, what is the role of the substrate, such as fast red TR?
What type of product is generated as an indicator of enzyme activity for both NADH diaphorase and SDH (Succinate Dehydrogenase)?
What type of product is generated as an indicator of enzyme activity for both NADH diaphorase and SDH (Succinate Dehydrogenase)?
How does the naphthol AS-D chloroacetate esterase stain differ from MOST other enzyme histochemical techniques?
How does the naphthol AS-D chloroacetate esterase stain differ from MOST other enzyme histochemical techniques?
Following an ATPase stain, a tissue section shows dark blue deposits at the sites of enzyme activity. What does this indicate?
Following an ATPase stain, a tissue section shows dark blue deposits at the sites of enzyme activity. What does this indicate?
The pH of the NADH diaphorase reaction is critical. What problem can arise if the pH deviates significantly from the optimum range?
The pH of the NADH diaphorase reaction is critical. What problem can arise if the pH deviates significantly from the optimum range?
A muscle biopsy shows a NEGATIVE phosphorylase reaction. What condition does this finding suggest?
A muscle biopsy shows a NEGATIVE phosphorylase reaction. What condition does this finding suggest?
Which of the following histochemical reactions involves the HYDROLYSIS of ester bonds to release a naphthol?
Which of the following histochemical reactions involves the HYDROLYSIS of ester bonds to release a naphthol?
What type of histochemical reaction demonstrates alkaline phosphatase activity?
What type of histochemical reaction demonstrates alkaline phosphatase activity?
Which ultrastructural criterion indicates good fixation in electron microscopy?
Which ultrastructural criterion indicates good fixation in electron microscopy?
What is a key advantage of using primary osmium fixation in electron microscopy?
What is a key advantage of using primary osmium fixation in electron microscopy?
A laboratory is considering osmium tetroxide for electron microscopy sample preparation. What is a significant disadvantage they should consider?
A laboratory is considering osmium tetroxide for electron microscopy sample preparation. What is a significant disadvantage they should consider?
What is a notable advantage of using aldehyde fixatives, like formaldehyde and glutaraldehyde, in primary fixation?
What is a notable advantage of using aldehyde fixatives, like formaldehyde and glutaraldehyde, in primary fixation?
What is a potential disadvantage of using aldehyde fixatives in immunohistochemistry?
What is a potential disadvantage of using aldehyde fixatives in immunohistochemistry?
What is a key advantage of using primary buffered picric acid-formaldehyde (PAF) fixative?
What is a key advantage of using primary buffered picric acid-formaldehyde (PAF) fixative?
A laboratory is considering using PAF as a fixative. What is a primary safety concern they must address?
A laboratory is considering using PAF as a fixative. What is a primary safety concern they must address?
What defines a dual-purpose fixative regarding tissue preservation?
What defines a dual-purpose fixative regarding tissue preservation?
Why is it necessary to heat and alkalinize paraformaldehyde when preparing fixative solutions?
Why is it necessary to heat and alkalinize paraformaldehyde when preparing fixative solutions?
What is the recommended maximum duration for tissues to remain in osmium tetroxide to avoid harm?
What is the recommended maximum duration for tissues to remain in osmium tetroxide to avoid harm?
How long can tissues generally remain in formaldehyde without significant harm?
How long can tissues generally remain in formaldehyde without significant harm?
Which of the following accurately compares epoxy resin embedding with paraffin embedding for tissue processing?
Which of the following accurately compares epoxy resin embedding with paraffin embedding for tissue processing?
What is the primary function of a transitional solvent in tissue processing?
What is the primary function of a transitional solvent in tissue processing?
What influence does the pH of a fixative solution have on fixation quality for electron microscopy?
What influence does the pH of a fixative solution have on fixation quality for electron microscopy?
What is the role of Dodecenyl succinic anhydride (DDSA) in Epon and Araldite embedding media?
What is the role of Dodecenyl succinic anhydride (DDSA) in Epon and Araldite embedding media?
Which measure is important for facilitating easier sectioning of resin-embedded tissue?
Which measure is important for facilitating easier sectioning of resin-embedded tissue?
What type of knife is preferred for cutting ultra-thin sections for electron microscopy?
What type of knife is preferred for cutting ultra-thin sections for electron microscopy?
What is the main purpose of cutting 0.5-µm sections of resin-embedded tissue?
What is the main purpose of cutting 0.5-µm sections of resin-embedded tissue?
If sections appear wrinkled during sectioning of resin-embedded tissue, what is the MOST appropriate corrective action?
If sections appear wrinkled during sectioning of resin-embedded tissue, what is the MOST appropriate corrective action?
How does section thickness affect the color intensity of stained tissue sections?
How does section thickness affect the color intensity of stained tissue sections?
What is the preferred thickness for thin sections intended for electron microscopy?
What is the preferred thickness for thin sections intended for electron microscopy?
What is the purpose of using Toluidine blue to stain 0.5-µm plastic sections?
What is the purpose of using Toluidine blue to stain 0.5-µm plastic sections?
Which heavy metal is commonly used for staining thin sections to enhance visibility of cellular structures under electron microscopy?
Which heavy metal is commonly used for staining thin sections to enhance visibility of cellular structures under electron microscopy?
Why is it important to exclude air and breath from lead citrate staining solution?
Why is it important to exclude air and breath from lead citrate staining solution?
What term describes areas within a sample that allow electrons to pass through more easily, appearing lighter in electron micrographs?
What term describes areas within a sample that allow electrons to pass through more easily, appearing lighter in electron micrographs?
Why should glass bottles be avoided when storing or using lead citrate solution?
Why should glass bottles be avoided when storing or using lead citrate solution?
Which cellular organelle contains cristae and produces energy?
Which cellular organelle contains cristae and produces energy?
Which of the following is NOT a step to make sectioning of resin-embedded tissue easier?
Which of the following is NOT a step to make sectioning of resin-embedded tissue easier?
Which of the following is NOT a corrective action for compression?
Which of the following is NOT a corrective action for compression?
If a tissue section shows poor differentiation between muscle and collagen. Which reagent is MOST likely to be adjusted to correct this?
If a tissue section shows poor differentiation between muscle and collagen. Which reagent is MOST likely to be adjusted to correct this?
What technique is MOST similar to the Bielschowsky stain in terms of the tissue components it highlights?
What technique is MOST similar to the Bielschowsky stain in terms of the tissue components it highlights?
Which ultrastructural criterion is MOST indicative of well-fixed tissue?
Which ultrastructural criterion is MOST indicative of well-fixed tissue?
In electron microscopy, uniform staining quality indicates good fixation. What does non-uniform staining MOST likely suggest?
In electron microscopy, uniform staining quality indicates good fixation. What does non-uniform staining MOST likely suggest?
Why is osmium tetroxide particularly effective in preserving cellular membranes for electron microscopy?
Why is osmium tetroxide particularly effective in preserving cellular membranes for electron microscopy?
Which of the following is a MAJOR limitation of using osmium tetroxide as a primary fixative?
Which of the following is a MAJOR limitation of using osmium tetroxide as a primary fixative?
Why are aldehyde fixatives, such as formaldehyde and glutaraldehyde, widely used in histology?
Why are aldehyde fixatives, such as formaldehyde and glutaraldehyde, widely used in histology?
Which of the following is a CONSIDERABLE drawback of using aldehyde fixatives in immunohistochemistry?
Which of the following is a CONSIDERABLE drawback of using aldehyde fixatives in immunohistochemistry?
What PRIMARY advantage does Buffered Picric Acid-Formaldehyde (PAF) fixative offer over formaldehyde alone?
What PRIMARY advantage does Buffered Picric Acid-Formaldehyde (PAF) fixative offer over formaldehyde alone?
Why is careful handling and storage EXTREMELY important when using picric acid-containing fixatives?
Why is careful handling and storage EXTREMELY important when using picric acid-containing fixatives?
Which BEST describes a dual-purpose fixative in the context of tissue preservation?
Which BEST describes a dual-purpose fixative in the context of tissue preservation?
Why is paraformaldehyde solution typically heated and alkalinized during preparation?
Why is paraformaldehyde solution typically heated and alkalinized during preparation?
What is the MOST likely consequence of leaving tissue in osmium tetroxide for an extended period?
What is the MOST likely consequence of leaving tissue in osmium tetroxide for an extended period?
What is the PRIMARY concern when tissues remain in formaldehyde for excessively long durations?
What is the PRIMARY concern when tissues remain in formaldehyde for excessively long durations?
What distinguishes epoxy resin embedding from paraffin embedding in tissue processing?
What distinguishes epoxy resin embedding from paraffin embedding in tissue processing?
What BEST describes the role of a transitional solvent in tissue processing?
What BEST describes the role of a transitional solvent in tissue processing?
How does the temperature at which fixation is performed influence the preservation of fine structures in electron microscopy?
How does the temperature at which fixation is performed influence the preservation of fine structures in electron microscopy?
What is the function of DMP-30 in Epon, Araldite, and Spurr embedding media?
What is the function of DMP-30 in Epon, Araldite, and Spurr embedding media?
What is the MOST effective way to minimize tearing during sectioning of resin-embedded tissue?
What is the MOST effective way to minimize tearing during sectioning of resin-embedded tissue?
What is the typical purpose of cutting 0.5-µm sections from resin-embedded tissue?
What is the typical purpose of cutting 0.5-µm sections from resin-embedded tissue?
What is the MOST appropriate corrective action if chatter marks are observed on sections?
What is the MOST appropriate corrective action if chatter marks are observed on sections?
Which of the following best describes the purpose of diagnostic cytology?
Which of the following best describes the purpose of diagnostic cytology?
What is the primary advantage of liquid-based cytology over traditional smear methods?
What is the primary advantage of liquid-based cytology over traditional smear methods?
Which of the following scenarios exemplifies cross contamination in a cytology lab?
Which of the following scenarios exemplifies cross contamination in a cytology lab?
Why is a cell block preparation particularly useful in diagnostic cytology?
Why is a cell block preparation particularly useful in diagnostic cytology?
Which of the following is an example of a gynecologic specimen?
Which of the following is an example of a gynecologic specimen?
For what purpose are nongynecologic specimens used in cytological examination?
For what purpose are nongynecologic specimens used in cytological examination?
What is the primary purpose of using a pre-fixative in cytology?
What is the primary purpose of using a pre-fixative in cytology?
How do alcohol-based pre-fixatives contribute to the quality of cytological preparations?
How do alcohol-based pre-fixatives contribute to the quality of cytological preparations?
What is the MOST significant effect of pre-fixatives on cellular morphology?
What is the MOST significant effect of pre-fixatives on cellular morphology?
Which fixative is LEAST suitable for preserving antigenicity for immunohistochemical staining in cell blocks?
Which fixative is LEAST suitable for preserving antigenicity for immunohistochemical staining in cell blocks?
For which type of sample is the direct smear method MOST appropriate?
For which type of sample is the direct smear method MOST appropriate?
What is the key advantage of using the cytocentrifuge method for sparsely cellular specimens?
What is the key advantage of using the cytocentrifuge method for sparsely cellular specimens?
In the ThinPrep method, how is a thin, uniform layer of cells achieved on the microscope slide?
In the ThinPrep method, how is a thin, uniform layer of cells achieved on the microscope slide?
What is the MOST effective method for handling a sparsely cellular specimen to ensure sufficient cells are available for examination?
What is the MOST effective method for handling a sparsely cellular specimen to ensure sufficient cells are available for examination?
Why might acetic acid be used in the preparation of a cytology smear?
Why might acetic acid be used in the preparation of a cytology smear?
What is the purpose of coating slides with poly-L-lysine or silane?
What is the purpose of coating slides with poly-L-lysine or silane?
Which of the following agents is BEST suited to remove excess mucus from a cytology sample?
Which of the following agents is BEST suited to remove excess mucus from a cytology sample?
What is the MOST likely consequence of failing to remove Carbowax fixatives from slides before staining?
What is the MOST likely consequence of failing to remove Carbowax fixatives from slides before staining?
What principle underlies the direct sedimentation method for making cell blocks?
What principle underlies the direct sedimentation method for making cell blocks?
Which cell block method is particularly advantageous for small cell samples due to its good structural integrity?
Which cell block method is particularly advantageous for small cell samples due to its good structural integrity?
Why is the Plasma-Thrombin method well-suited for cytology samples with low cellularity?
Why is the Plasma-Thrombin method well-suited for cytology samples with low cellularity?
A cytologist observes that a cell block is too fragile and falls apart during sectioning. Which cell block method would MOST likely prevent this issue in the future?
A cytologist observes that a cell block is too fragile and falls apart during sectioning. Which cell block method would MOST likely prevent this issue in the future?
What is the PRIMARY purpose of the Papanicolaou (Pap) stain?
What is the PRIMARY purpose of the Papanicolaou (Pap) stain?
Which of the following dyes stains cell nuclei blue in the Papanicolaou stain?
Which of the following dyes stains cell nuclei blue in the Papanicolaou stain?
What cellular component is specifically targeted by Orange G in the Papanicolaou staining technique?
What cellular component is specifically targeted by Orange G in the Papanicolaou staining technique?
What is the function of the EA component (Eosin Azure) in the Papanicolaou stain?
What is the function of the EA component (Eosin Azure) in the Papanicolaou stain?
What corrective action should be taken if a Pap-stained slide exhibits poor cell preservation and staining quality?
What corrective action should be taken if a Pap-stained slide exhibits poor cell preservation and staining quality?
A batch of Pap stains consistently shows overly intense nuclear staining. What is the MOST likely corrective action?
A batch of Pap stains consistently shows overly intense nuclear staining. What is the MOST likely corrective action?
In a cytology lab, what is the MOST effective approach for preventing cross-contamination between specimens?
In a cytology lab, what is the MOST effective approach for preventing cross-contamination between specimens?
What is the significance of using dedicated reagents for each specimen in a cytology laboratory?
What is the significance of using dedicated reagents for each specimen in a cytology laboratory?
A cytology technician notices cells from a previous sample on a staining rack. What immediate action should be taken to prevent cross-contamination?
A cytology technician notices cells from a previous sample on a staining rack. What immediate action should be taken to prevent cross-contamination?
What is the PRIMARY reason for clearly labeling and physically separating cytology specimens?
What is the PRIMARY reason for clearly labeling and physically separating cytology specimens?
Why is it important to provide proper training and standardized protocols to personnel in a cytology lab?
Why is it important to provide proper training and standardized protocols to personnel in a cytology lab?
Flashcards
Periodic Acid-Schiff (PAS)
Periodic Acid-Schiff (PAS)
Stains glycogen, mucopolysaccharides, and basement membranes magenta; nuclei stain blue.
PAS with Diastase
PAS with Diastase
Shows glycogen after digestion with diastase. Other carbohydrates stain magenta, nuclei stain blue.
Best Carmine
Best Carmine
Stains glycogen and mucins red.
Mayer Mucicarmine
Mayer Mucicarmine
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Alcian Blue, pH 2.5
Alcian Blue, pH 2.5
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Alcian Blue, pH 1.0
Alcian Blue, pH 1.0
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Müller-Mowry Colloidal Iron
Müller-Mowry Colloidal Iron
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Alcian Blue-PAS
Alcian Blue-PAS
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Congo Red
Congo Red
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Crystal Violet
Crystal Violet
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Thioflavin T
Thioflavin T
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Carbohydrate
Carbohydrate
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Polysaccharide
Polysaccharide
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Acid mucopolysaccharides
Acid mucopolysaccharides
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Birefringence
Birefringence
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Metachromasia
Metachromasia
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Polychromasia
Polychromasia
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Epithelial Mucins
Epithelial Mucins
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Connective Tissue Mucins
Connective Tissue Mucins
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Collagen fibers
Collagen fibers
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Elastic fibers
Elastic fibers
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Reticular fibers
Reticular fibers
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Skeletal muscle fibers
Skeletal muscle fibers
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Cardiac muscle fibers
Cardiac muscle fibers
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Smooth muscle fibers
Smooth muscle fibers
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Fibroblasts
Fibroblasts
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Macrophages
Macrophages
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Mast cells
Mast cells
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Adipocytes
Adipocytes
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Chondrocytes
Chondrocytes
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Masson trichrome
Masson trichrome
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Gomori 1-step trichrome
Gomori 1-step trichrome
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Van Gieson
Van Gieson
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Verhoeff
Verhoeff
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Gomori aldehyde fuchsin
Gomori aldehyde fuchsin
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Orcein
Orcein
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Gomori silver impregnation
Gomori silver impregnation
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Gordon and Sweets silver impregnation
Gordon and Sweets silver impregnation
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Methenamine-silver
Methenamine-silver
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Mallory PTAH
Mallory PTAH
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Oil red O
Oil red O
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Sudan black B
Sudan black B
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Osmium tetroxide
Osmium tetroxide
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Toluidine blue
Toluidine blue
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Methyl green-pyronin
Methyl green-pyronin
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Masson trichrome use
Masson trichrome use
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Gomori 1-step trichrome use
Gomori 1-step trichrome use
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Van Gieson use
Van Gieson use
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Verhoeff use
Verhoeff use
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Gomori aldehyde fuchsin use
Gomori aldehyde fuchsin use
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Orcein use
Orcein use
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Methenamine-silver use
Methenamine-silver use
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Mallory PTAH use
Mallory PTAH use
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Oil red O use
Oil red O use
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Sudan black B use
Sudan black B use
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Methyl green-pyronin use
Methyl green-pyronin use
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Oil Red O technique
Oil Red O technique
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Osmium Tetroxide technique
Osmium Tetroxide technique
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Sudan Black B technique
Sudan Black B technique
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Mast cell feature
Mast cell feature
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Pyronin target
Pyronin target
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Impregnation
Impregnation
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Neuron
Neuron
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Glia and Glial Fibers
Glia and Glial Fibers
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Nissl Substance
Nissl Substance
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Myelin
Myelin
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Axon (Axis Cylinder)
Axon (Axis Cylinder)
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Cresyl Echt Violet
Cresyl Echt Violet
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Bodian Stain
Bodian Stain
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Holmes Silver Nitrate
Holmes Silver Nitrate
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Bielschowsky Stain
Bielschowsky Stain
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Sevier-Munger Stain
Sevier-Munger Stain
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Phosphotungstic Acid-Hematoxylin (PTAH)
Phosphotungstic Acid-Hematoxylin (PTAH)
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Holzer Stain
Holzer Stain
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Cajal Stain
Cajal Stain
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Weil Stain
Weil Stain
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Luxol Fast Blue
Luxol Fast Blue
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Bacteria
Bacteria
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Cocci
Cocci
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Bacilli
Bacilli
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Spirochetes
Spirochetes
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Mycobacteria
Mycobacteria
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Fungi
Fungi
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Yeasts
Yeasts
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Protozoans
Protozoans
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Hyphae
Hyphae
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Acid-Fast Techniques
Acid-Fast Techniques
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Auramine-Rhodamine
Auramine-Rhodamine
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Gram Stain Modifications
Gram Stain Modifications
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Giemsa
Giemsa
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Alcian Yellow-Toluidine Blue
Alcian Yellow-Toluidine Blue
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Chromic Acid-Schiff (CAS)
Chromic Acid-Schiff (CAS)
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Gridley Fungus Stain
Gridley Fungus Stain
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Grocott Modification of Gomori Methenamine-Silver
Grocott Modification of Gomori Methenamine-Silver
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Warthin-Starry
Warthin-Starry
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Dieterle
Dieterle
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Steiner and Steiner
Steiner and Steiner
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10% Neutral Buffered Formalin
10% Neutral Buffered Formalin
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Acid-alcohol in Acid-Fast Staining
Acid-alcohol in Acid-Fast Staining
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Methylene Blue or Malachite Green
Methylene Blue or Malachite Green
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Acid-Fast Control Material
Acid-Fast Control Material
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Error in Acid-Fast Staining
Error in Acid-Fast Staining
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Mode of Action in Acid-Fast
Mode of Action in Acid-Fast
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Chemically Clean Glassware
Chemically Clean Glassware
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Light Microscope with Oil Immersion
Light Microscope with Oil Immersion
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Rhodamine B
Rhodamine B
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Methanol or Heat Fixation
Methanol or Heat Fixation
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Iodine (Mordant)
Iodine (Mordant)
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Endogenous Pigment
Endogenous Pigment
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Exogenous Pigment
Exogenous Pigment
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Hematogenous Pigment
Hematogenous Pigment
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Anthracotic Pigment
Anthracotic Pigment
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Endogenous, Nonhematogenous Pigment
Endogenous, Nonhematogenous Pigment
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Mineral
Mineral
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Argentaffin Reaction
Argentaffin Reaction
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Argyrophil Reaction
Argyrophil Reaction
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Argentaffin & Argyrophil Similarities
Argentaffin & Argyrophil Similarities
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Argentaffin & Argyrophil Differences
Argentaffin & Argyrophil Differences
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Prussian Blue
Prussian Blue
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Turnbull Blue
Turnbull Blue
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Schmorl Ferric-Ferricyanide Reduction Test
Schmorl Ferric-Ferricyanide Reduction Test
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Fontana-Masson
Fontana-Masson
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Grimelius
Grimelius
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Churukian-Schenk
Churukian-Schenk
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Gomori Methenamine-Silver
Gomori Methenamine-Silver
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Hall (Fouchet)
Hall (Fouchet)
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von Kossa
von Kossa
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Alizarin Red S
Alizarin Red S
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Rhodanine
Rhodanine
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Prussian Blue/Turnbull Blue Fixative
Prussian Blue/Turnbull Blue Fixative
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Prussian Blue Reagents
Prussian Blue Reagents
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Prussian Blue Result
Prussian Blue Result
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Prussian Blue Control
Prussian Blue Control
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Turnbull Blue Reagents
Turnbull Blue Reagents
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Turnbull Blue Result
Turnbull Blue Result
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Turnbull Blue Control
Turnbull Blue Control
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Schmorl Fixative
Schmorl Fixative
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Schmorl Reagents
Schmorl Reagents
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Schmorl Result
Schmorl Result
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Schmorl Control
Schmorl Control
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Fontana-Masson Reagent
Fontana-Masson Reagent
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Fontana-Masson Result
Fontana-Masson Result
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Fontana-Masson Control
Fontana-Masson Control
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Grimelius Reagent
Grimelius Reagent
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Grimelius Result
Grimelius Result
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Grimelius Control
Grimelius Control
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Churukian-Schenk Reagent
Churukian-Schenk Reagent
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Churukian-Schenk Result
Churukian-Schenk Result
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Churukian-Schenk Control
Churukian-Schenk Control
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GMS Reagents
GMS Reagents
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GMS Result
GMS Result
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Hall's Stain Control Tissue
Hall's Stain Control Tissue
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von Kossa Reagents
von Kossa Reagents
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von Kossa Result
von Kossa Result
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von Kossa Control Tissue
von Kossa Control Tissue
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Melanin Locations
Melanin Locations
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Anthracotic Pigment Test
Anthracotic Pigment Test
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Formalin Pigment Test
Formalin Pigment Test
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Melanin Pigment Test
Melanin Pigment Test
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Antigen
Antigen
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Antibody
Antibody
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Substrate
Substrate
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Chromogen
Chromogen
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Fluorochrome
Fluorochrome
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Epitope
Epitope
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Secondary Antibody
Secondary Antibody
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Multilink Secondary Antiserum
Multilink Secondary Antiserum
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Polyclonal Antisera
Polyclonal Antisera
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Monoclonal Antibodies
Monoclonal Antibodies
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Direct Immunohistochemical Technique
Direct Immunohistochemical Technique
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Indirect Immunohistochemical Technique
Indirect Immunohistochemical Technique
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Peroxidase-Antiperoxidase (PAP)
Peroxidase-Antiperoxidase (PAP)
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Avidin-Biotin Complex (ABC)
Avidin-Biotin Complex (ABC)
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Polymeric Detection System
Polymeric Detection System
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Fluorescein Isothiocyanate (FITC)
Fluorescein Isothiocyanate (FITC)
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3,3'-Diaminobenzidine (DAB)
3,3'-Diaminobenzidine (DAB)
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4-Chloro-1-naphthol (4-CN)
4-Chloro-1-naphthol (4-CN)
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Fast Red
Fast Red
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New Fuchsin
New Fuchsin
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Peroxidase Enzyme System
Peroxidase Enzyme System
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Alkaline Phosphatase Enzyme System
Alkaline Phosphatase Enzyme System
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Specimen Prep for Immunofluorescence
Specimen Prep for Immunofluorescence
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Problems with Formaldehyde Fixation
Problems with Formaldehyde Fixation
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Glutaraldehyde Special Issues
Glutaraldehyde Special Issues
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Alcohol-based Fixatives Special Issues
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Bouin's Solution Special Issues
Bouin's Solution Special Issues
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Zinc-based Fixatives Special Issues
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Protein Blocking
Protein Blocking
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Endogenous Peroxidase Blocking
Endogenous Peroxidase Blocking
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5 Heavy Chains of Immunoglobulins
5 Heavy Chains of Immunoglobulins
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2 Light Chains of Immunoglobulins
2 Light Chains of Immunoglobulins
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Negative Tissue Controls Purpose
Negative Tissue Controls Purpose
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Negative Reagent Controls Purpose
Negative Reagent Controls Purpose
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Corrective Action for Non-specific Binding
Corrective Action for Non-specific Binding
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Corrective Action for Weak/No Staining
Corrective Action for Weak/No Staining
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Corrective Action for High Background Staining
Corrective Action for High Background Staining
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Corrective Action for Over-staining
Corrective Action for Over-staining
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Corrective Action for Autofluorescence
Corrective Action for Autofluorescence
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Corrective Action for Edge Artifact
Corrective Action for Edge Artifact
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Corrective Action for Inconsistent Staining
Corrective Action for Inconsistent Staining
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Advantages of Preparing Positive Controls
Advantages of Preparing Positive Controls
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Epitope Retrieval Methods
Epitope Retrieval Methods
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Solutions Used for HIER
Solutions Used for HIER
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Ways of Heating in HIER
Ways of Heating in HIER
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Solutions Used for EIER
Solutions Used for EIER
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Alcohol-Soluble Chromogens
Alcohol-Soluble Chromogens
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Non-Alcohol-Soluble Chromogens
Non-Alcohol-Soluble Chromogens
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Chemicals Intensifying DAB Reaction
Chemicals Intensifying DAB Reaction
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Enzymes
Enzymes
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Oxidation
Oxidation
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Reduction
Reduction
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Hydrolase
Hydrolase
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Simultaneous Coupling
Simultaneous Coupling
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Esterase
Esterase
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Phosphatase
Phosphatase
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Phosphorylase
Phosphorylase
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Epimysium
Epimysium
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Perimysium
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Endomysium
Endomysium
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Skeletal Muscle
Skeletal Muscle
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Cardiac Muscle
Cardiac Muscle
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Smooth Muscle
Smooth Muscle
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Type I Muscle Fibers
Type I Muscle Fibers
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Type II Muscle Fibers
Type II Muscle Fibers
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Type IIa Muscle Fibers
Type IIa Muscle Fibers
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Type IIb Muscle Fibers
Type IIb Muscle Fibers
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Enzyme Specificity
Enzyme Specificity
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Catalytic Efficiency
Catalytic Efficiency
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Enzyme Regulation
Enzyme Regulation
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pH Influence on Enzymes
pH Influence on Enzymes
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Temperature Influence on Enzymes
Temperature Influence on Enzymes
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Substrate Concentration Influence
Substrate Concentration Influence
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Cofactors and Coenzymes
Cofactors and Coenzymes
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Inhibitors and Activators
Inhibitors and Activators
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Freezing Artifact
Freezing Artifact
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Cryoprotective Solution
Cryoprotective Solution
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Snap-Freezing
Snap-Freezing
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α-Naphthyl Acetate Esterase Reaction
α-Naphthyl Acetate Esterase Reaction
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Naphthol AS-D Chloroacetate Esterase Reaction
Naphthol AS-D Chloroacetate Esterase Reaction
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ATPase Reaction
ATPase Reaction
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Acid Phosphatase Reaction
Acid Phosphatase Reaction
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Alkaline Phosphatase Reaction
Alkaline Phosphatase Reaction
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NADH Diaphorase Reaction
NADH Diaphorase Reaction
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SDH (Succinate Dehydrogenase) Reaction
SDH (Succinate Dehydrogenase) Reaction
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Phosphorylase Reaction
Phosphorylase Reaction
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Naphthol AS-D Chloroacetate Esterase Uniqueness
Naphthol AS-D Chloroacetate Esterase Uniqueness
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McArdle Disease
McArdle Disease
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Cellular & Organelle Integrity
Cellular & Organelle Integrity
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Absence of Artifacts
Absence of Artifacts
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Clear and Distinct Membranes
Clear and Distinct Membranes
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Cytoplasmic Matrix Preservation
Cytoplasmic Matrix Preservation
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Retention of Extracellular Matrix
Retention of Extracellular Matrix
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Staining Quality
Staining Quality
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Osmium: Membrane Preservation
Osmium: Membrane Preservation
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Osmium: Enhanced TEM Contrast
Osmium: Enhanced TEM Contrast
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Osmium: Toxicity
Osmium: Toxicity
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Osmium: Limited Penetration
Osmium: Limited Penetration
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Osmium: High Cost
Osmium: High Cost
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Aldehydes: Wide Applicability
Aldehydes: Wide Applicability
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Aldehydes: Good Penetration
Aldehydes: Good Penetration
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Aldehydes: Preserve Fine Structure
Aldehydes: Preserve Fine Structure
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Aldehydes: Versatility
Aldehydes: Versatility
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Aldehydes: Stable Fixation
Aldehydes: Stable Fixation
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Aldehydes: Staining Compatibility
Aldehydes: Staining Compatibility
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Aldehydes: Protein Cross-Linking
Aldehydes: Protein Cross-Linking
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Aldehydes: Slow Penetration
Aldehydes: Slow Penetration
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Aldehydes: Potential for Toxicity
Aldehydes: Potential for Toxicity
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PAF: Morphological Preservation
PAF: Morphological Preservation
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PAF: Good Penetration
PAF: Good Penetration
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PAF: Staining Techniques
PAF: Staining Techniques
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PAF: Antigen Masking
PAF: Antigen Masking
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PAF: Picric Acid Explosiveness
PAF: Picric Acid Explosiveness
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PAF: Yellow Pigmentation
PAF: Yellow Pigmentation
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Dual-Purpose Fixative
Dual-Purpose Fixative
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Heating Paraformaldehyde
Heating Paraformaldehyde
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Alkalinizing Paraformaldehyde
Alkalinizing Paraformaldehyde
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Osmium Tetroxide Duration
Osmium Tetroxide Duration
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Glutaraldehyde Duration
Glutaraldehyde Duration
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Formaldehyde Duration
Formaldehyde Duration
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PAF Duration
PAF Duration
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Formaldehyde-Glutaraldehyde Duration
Formaldehyde-Glutaraldehyde Duration
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Epoxy Resin Fixation
Epoxy Resin Fixation
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Paraffin Embedding Fixation
Paraffin Embedding Fixation
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Epoxy & Paraffin Dehydration
Epoxy & Paraffin Dehydration
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Epoxy Transitional Solvent
Epoxy Transitional Solvent
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Paraffin Transitional Solvent
Paraffin Transitional Solvent
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Epoxy Sectioning
Epoxy Sectioning
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Paraffin Sectioning
Paraffin Sectioning
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Purpose of Epoxy Embedding
Purpose of Epoxy Embedding
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Purpose of Paraffin Embedding
Purpose of Paraffin Embedding
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Resolution of Epoxy
Resolution of Epoxy
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Transitional Solvent
Transitional Solvent
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Fixative Type Influence
Fixative Type Influence
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Concentration Influence
Concentration Influence
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Temperature Influence
Temperature Influence
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pH Influence
pH Influence
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Osmolality Influence
Osmolality Influence
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Penetration Rate Influence
Penetration Rate Influence
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Epon/Araldite/Spurr Resin Function
Epon/Araldite/Spurr Resin Function
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DDSA Function
DDSA Function
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MNA Function
MNA Function
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DMP-30 Function
DMP-30 Function
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Spurr Resin
Spurr Resin
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Trimming resins
Trimming resins
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Block Orientation
Block Orientation
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Diagnostic Cytology
Diagnostic Cytology
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Liquid-Based Cytology
Liquid-Based Cytology
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Cross Contamination
Cross Contamination
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Cell Block
Cell Block
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Gynecologic Specimens
Gynecologic Specimens
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Nongynecologic Specimens
Nongynecologic Specimens
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Use of Pre-fixatives
Use of Pre-fixatives
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Effect of Pre-fixatives on Cellular Morphology
Effect of Pre-fixatives on Cellular Morphology
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95% Ethanol
95% Ethanol
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Direct Smear Method
Direct Smear Method
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Cytocentrifuge Method
Cytocentrifuge Method
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ThinPrep Method
ThinPrep Method
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Handle Sparsely Cellular Specimens
Handle Sparsely Cellular Specimens
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Reduce Red Blood Cells in Smears
Reduce Red Blood Cells in Smears
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Increase Cellular Adherence to Slides
Increase Cellular Adherence to Slides
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Remove Excess Mucus
Remove Excess Mucus
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Staining Effect: Carbowax Not Removed
Staining Effect: Carbowax Not Removed
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Cell Block: Direct Sedimentation
Cell Block: Direct Sedimentation
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Cell Block: Agar Method
Cell Block: Agar Method
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Cell Block: Plasma-Thrombin Method
Cell Block: Plasma-Thrombin Method
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Cell Block: Gelatin Method
Cell Block: Gelatin Method
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Pap Stain Purpose
Pap Stain Purpose
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Hematoxylin
Hematoxylin
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Orange G
Orange G
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EA (Eosin Azure)
EA (Eosin Azure)
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Error: Inadequate Fixation
Error: Inadequate Fixation
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Corrective Action: Over/Under staining
Corrective Action: Over/Under staining
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Error: Contamination of Staining Reagents
Error: Contamination of Staining Reagents
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Corrective Action: Poor Slide Preparation
Corrective Action: Poor Slide Preparation
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Cross-Contamination Occurrence
Cross-Contamination Occurrence
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Techniques to Prevent Cross-Contamination
Techniques to Prevent Cross-Contamination
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Primary Reagents and/or Dyes
Primary Reagents and/or Dyes
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Study Notes
- A neuron is a specialized cell transmitting nerve impulses
- Glia are supportive cells in the central nervous system that do not conduct electrical impulses
- Glial fibers are extensions from glial cells
- Nissl substance is a granular body in neurons consisting of rough endoplasmic reticulum and ribosomes, used for protein synthesis
- Myelin is an insulating layer around nerves, made of protein and fatty substances
- An axon (axis cylinder) is the long threadlike part of a nerve cell that impulses are conducted along
Techniques Classified by Element Demonstrated
- Cresyl echt violet stains Nissl substance (rough endoplasmic reticulum in neurons)
- Bodian stains nerve fibers and neurofibrils
- Holmes silver nitrate stains nerve fibers
- Bielschowsky stains nerve fibers and neurofibrils
- Sevier-Munger stains nerve fibers and neurofibrils
- Thioflavin S stains amyloid
- Phosphotungstic acid-hematoxylin (PTAH) stains muscle striations, fibrin, and glial fibers
- Holzer stains glial fibers
- Cajal stain stains astrocytes
- Weil stains myelin
- Luxol fast blue stains myelin
- Acid-fast techniques (Kinyoun, Ziehl-Neelsen, Fite): Mycobacteria (e.g., Mycobacterium tuberculosis)
- Auramine-rhodamine: Mycobacteria (e.g., Mycobacterium tuberculosis)
- Gram stain modifications: Bacteria (differentiates between Gram-positive and Gram-negative bacteria)
- Giemsa: Protozoans and bacteria (e.g., Plasmodium species, Helicobacter pylori)
- Alcian yellow-toluidine blue: Helicobacter pylori (bacteria)
- Periodic acid-Schiff (PAS): Fungi and certain bacteria (e.g., Candida species, Tropheryma whipplei)
- Chromic acid-Schiff (CAS): Fungi (e.g., Histoplasma species)
- Gridley fungus stain: Fungi (e.g., Aspergillus species)
- Grocott modification of Gomori methenamine-silver: Fungi (e.g., Pneumocystis jirovecii)
- Mayer mucicarmine: Fungi and Cryptococcus species (bacteria)
- Warthin-Starry: Spirochetes (e.g., Treponema pallidum)
- Dieterle: Spirochetes (e.g., Legionella species)
- Steiner and Steiner: Bacteria, including spirochetes and Helicobacter pylori
- Prussian blue demonstrates ferric iron (Fe³⁺)
- Turnbull blue demonstrates ferrous iron (Fe²⁺)
- Schmorl ferric-ferricyanide reduction demonstrates reducing substances like melanin, lipofuscin, and argentaffin cells
- Fontana-Masson demonstrates melanin and argentaffin granules
- Grimelius demonstrates argyrophil granules
- Churukian-Schenk demonstrates argyrophil granules
- Gomori methenamine-silver demonstrates basement membranes and fungi
- Hall (Fouchet) demonstrates bilirubin
- Von Kossa demonstrates calcium
- Alizarin red S demonstrates calcium
- Rhodanine demonstrates copper
Cresyl Echt Violet Stain Details
- Formalin is the most desirable fixative
- Refixation in formalin may be necessary if another fixative has been used
- Cresyl echt violet stains Nissl substance (rough endoplasmic reticulum) in neurons
- Nissl bodies in neurons appear violet after staining
- Brain tissue known to have Nissl substance is appropriate control material
- Adjust staining time to correct overstaining or understaining problems
- Cresyl echt violet binds to RNA in the Nissl substance
- Use chemically clean glassware
- Uses light microscopy
Bodian Stain Details
- Formalin is the most desirable fixative
- Refixation in formalin may be necessary if another fixative has been used
- Protargol (silver protein) stains nerve fibers and neurofibrils
- Nerve fibers and neurofibrils appear black after staining
- Nervous tissue known to have nerve fibers and neurofibrils is appropriate control material
- Ensure proper timing and washing steps to fix incomplete impregnation or overstaining
- A silver impregnation technique where silver binds to neurofibrils
- Use chemically clean glassware
- Uses light microscopy
Holmes Silver Nitrate Stain Details
- Formalin is the most desirable fixative
- Refixation in formalin may be necessary if another fixative has been used
- Silver nitrate stains nerve fibers and neurofibrils
- Nerve fibers and neurofibrils appear black after staining
- Nervous tissue known to have nerve fibers and neurofibrils is appropriate control material
- Ensure proper timing and washing steps to fix incomplete impregnation or overstaining
- A silver impregnation technique where silver binds to nerve fibers and neurofibrils
- Use chemically clean glassware
- Uses light microscopy
Bielschowsky Stain Details
- Formalin is the most desirable fixative
- Refixation in formalin may be necessary if another fixative has been used
- Silver nitrate stains nerve fibers and neurofibrils
- Nerve fibers and neurofibrils appear black after staining
- Nervous tissue known to have nerve fibers and neurofibrils is appropriate control material
- Ensure proper timing and washing steps to fix incomplete impregnation or overstaining
- A silver impregnation technique where silver binds to nerve fibers and neurofibrils
- Use chemically clean glassware
- Uses light microscopy
Sevier-Munger Stain Details
- Formalin is the most desirable fixative
- Refixation in formalin may be necessary if another fixative has been used
- Silver nitrate stains nerve fibers and neurofibrils
- Nerve fibers and neurofibrils appear black after staining
- Nervous tissue known to have nerve fibers and neurofibrils is appropriate control material
- Ensure proper timing and washing steps to fix incomplete impregnation or overstaining
- A silver impregnation technique where silver binds to nerve fibers and neurofibrils
- Use chemically clean glassware
- Uses light microscopy
Thioflavin S Stain Details
- Formalin is the most desirable fixative
- Refixation in formalin may be necessary if another fixative has been used
- Thioflavin S is a fluorescent dye used to stain amyloid
- Amyloid deposits will fluoresce green under UV light
- Tissue known to contain amyloid deposits is appropriate control material
- Adjust staining time and avoid prolonged exposure to light to correct overstaining or photobleaching
- Thioflavin S binds to beta-pleated sheet structures in amyloid fibrils, causing them to fluoresce
- Requires a UV light source for visualization
- Uses fluorescence microscopy
Phosphotungstic Acid-Hematoxylin (PTAH) Stain Details
- 10% neutral buffered formalin is the most desirable fixative
- The tissue can be post-fixed in formalin or Bouin's solution if another fixative has been used
- PTAH solution stains muscle striations, fibrin, and glial fibers blue, and collagen red-brown
- Muscle striations, fibrin, and glial fibers appear blue, while collagen appears red-brown
- Tissue sections containing muscle, fibrin, and glial fibers are appropriate control material
- Ensure proper timing and concentration of reagents to correct overstaining or understaining
- PTAH binds to tissue components based on their affinity for phosphotungstic acid and hematoxylin
- Use chemically clean glassware
- Uses light microscopy
Holzer Stain Details
- 10% neutral buffered formalin is the most desirable fixative
- The tissue can be post-fixed in formalin if another fixative has been used
- Crystal violet stains glial fibers; phosphomolybdic acid differentiates the staining
- Glial fibers appear blue or purple
- Brain tissue sections containing glial fibers are appropriate control material
- Ensure proper timing and concentration of reagents can occur to fix overstaining or understaining
- Crystal violet binds to glial fibers, and phosphomolybdic acid differentiates the staining
- Use chemically clean glassware
- Uses light microscopy
Cajal Stain Details
- Formalin or ammonium bromide are the most desirable fixatives
- Refixation in formalin or ammonium bromide may be necessary if another fixative has been used
- Gold sublimate stains astrocytes, formalin is used for fixation
- Astrocytes appear black against a yellow or light brown background
- Brain tissue known to contain astrocytes is appropriate control material
- Adjust staining time or concentration of reagents to correct overstaining or understaining
- Impregnation of astrocytes with gold sublimate
- Use chemically clean glassware
- Uses light microscopy
Weil Stain Details
- 10% formalin is the most desirable fixative
- Refixation in 10% formalin may be necessary if another fixative has been used
- Hematoxylin stains myelin sheaths, ferric ammonium sulfate is a mordant, borax ferricyanide is a differentiator
- Myelin sheaths appear blue-black, and the background is colorless or pale yellow
- Spinal cord or brain tissue known to contain myelin is appropriate control material
- Adjust differentiation time to correct overdifferentiation which can remove the stain from myelin
- Staining of myelin sheaths with hematoxylin and differentiation with ferric ammonium sulfate and borax ferricyanide
- Use chemically clean glassware
- Uses light microscopy
Luxol Fast Blue Stain Details
- 10% formalin is the most desirable fixative
- Refixation in 10% formalin may be necessary if another fixative has been used
- Luxol fast blue stains myelin, cresyl echt violet counterstains Nissl substance
- Myelin appears blue, and Nissl substance appears violet
- Brain or spinal cord tissue known to contain myelin and Nissl substance is appropriate control material
- Adjust staining time or concentration of reagents to correct overstaining or understaining
- Staining of myelin with Luxol fast blue and counterstaining of Nissl substance with cresyl echt violet
- Use chemically clean glassware
- Uses light microscopy
Definitions of Microorganisms
- Bacteria are single-celled microorganisms which can be free-living or parasitic
- Examples include Escherichia coli and Streptococcus pneumoniae
- Cocci are spherical-shaped bacteria
- Examples include Staphylococcus aureus and Streptococcus pyogenes
- Bacilli are rod-shaped bacteria
- Examples include Bacillus anthracis and Mycobacterium tuberculosis
- Spirochetes are spiral-shaped bacteria
- Examples include Treponema pallidum (causes syphilis) and Borrelia burgdorferi (causes Lyme disease)
- Mycobacteria is a genus of bacteria that includes pathogens known to cause serious diseases in mammals
- Examples include Mycobacterium tuberculosis (causes tuberculosis) and Mycobacterium leprae (causes leprosy)
- Fungi are a group of eukaryotic organisms which includes microorganisms such as yeasts and molds, as well as the more familiar mushrooms
- Examples include Aspergillus and Penicillium
- Yeasts are a type of fungus that is single-celled and can ferment carbohydrates
- Examples include Saccharomyces cerevisiae (brewer's yeast) and Candida albicans (can cause infections in humans)
- Protozoans are single-celled eukaryotes that can be free-living or parasitic
- Examples include Plasmodium (causes malaria) and Giardia lamblia (causes giardiasis)
- Hyphae are the long, branching filamentous structures of a fungus
- Examples include molds like Rhizopus and Aspergillus
- Mycelia is the vegetative part of a fungus, consisting of a network of fine white filaments (hyphae)
- Examples include mushrooms and molds
Acid-Fast Techniques (Kinyoun, Ziehl-Neelsen, Fite) Stain Details
- This stains Mycobacteria, for example, Mycobacterium tuberculosis.
- 10% neutral buffered formalin is the most desirable fixative.
- Tissue should be washed thoroughly if another fixative has been used to remove any residual fixative before staining.
- Carbol fuchsin stains acid-fast bacilli.
- Acid-alcohol decolorizes, removing the primary stain from non-acid-fast cells.
- Methylene blue or malachite green counterstains the background.
- Acid-fast bacilli appear red, and the background appears blue or green after staining.
- Tissue containing Mycobacterium tuberculosis is appropriate control material.
- Over-decolorization can lead to false negatives, so ensure proper timing and concentration of decolorizer.
- Mycolic acid in the cell walls of mycobacteria retains the carbol fuchsin dye even after decolorization with acid-alcohol.
- Use chemically clean glassware to avoid contamination.
- Uses a light microscope with an oil immersion lens.
Auramine-Rhodamine Stain Details
- This stains Mycobacteria, for example, Mycobacterium tuberculosis
- 10% neutral buffered formalin is the most desirable fixative
- Tissue should be washed thoroughly if another fixative has been used to remove any residual fixative before staining
- Auramine O is the primary stain, a fluorescent dye that binds to mycobacteria
- Rhodamine B is a secondary stain that enhances fluorescence
- Acid-alcohol decolorizes and removes non-specific staining
- Potassium permanganate is a counterstain which reduces background fluorescence
- Mycobacteria fluoresce yellow or orange under UV light after staining
- Tissue known to contain Mycobacterium tuberculosis is appropriate control material
- Over-decolorization can lead to false negatives; ensure proper timing and concentration of decolorizer
- The fluorescent dyes bind to the mycolic acids in the cell walls of mycobacteria
- Use chemically clean glassware to avoid contamination
- Uses a fluorescence microscope
Gram Stain Modifications Stain Details
- This stains stains bacteria, differentiating between Gram-positive and Gram-negative bacteria.
- Methanol or heat fixation is the most desirable fixative.
- Ensure the sample is adequately fixed to prevent washing off during staining if another fixative has been used.
- Crystal violet is the primary stain, which stains all cells.
- Iodine is a mordant and forms a complex with crystal violet.
- Alcohol or acetone is a decolorizer which removes the primary stain from Gram-negative cells.
- Safranin counterstains Gram-negative cells.
- Gram-positive bacteria appear purple, and Gram-negative bacteria appear pink/red after staining.
- Known Gram-positive and Gram-negative bacterial cultures make appropriate control material.
- Over-decolorization or under-decolorization can be a source of error, so ensure proper timing and technique during the decolorization step.
- Crystal violet stains all cells, iodine forms a complex with crystal violet, alcohol or acetone decolorizes Gram-negative cells, and safranin counterstains Gram-negative cells.
- Use chemically clean glassware to avoid contamination.
- Uses a light microscope.
Giemsa Stain Details
- Stains protozoans and bacteria (e.g., Plasmodium species, Helicobacter pylori)
- Methanol is the most desirable fixative
- Ensure the sample is adequately fixed to prevent washing off during staining if another fixative has been used
- Giemsa stain (a mixture of eosin and methylene blue) stains nucleic acids and cytoplasmic components
- Protozoans and bacteria are stained in shades of blue to purple, with the background appearing pink after staining
- Known samples containing the target protozoans or bacteria is appropriate control material
- Ensure proper fixation and follow staining protocol accurately; inadequate fixation or staining time can be a source of error
- Giemsa stain binds to nucleic acids and cytoplasmic components, differentiating cellular structures
- Use chemically clean glassware to avoid contamination
- Uses a light microscope
Alcian Yellow-Toluidine Blue Stain Details
- This stains Helicobacter pylori (bacteria)
- Formalin is the most desirable fixative
- Ensure the tissue is adequately washed and processed to remove any residual fixative that may interfere with staining if another fixative has been used
- Alcian yellow stains Helicobacter pylori yellow
- Toluidine blue: Stains the background tissue blue
- Helicobacter pylori bacteria appear yellow against a blue background after staining
- Tissue known to be infected with Helicobacter pylori is appropriate control material
- Ensure proper fixation and follow staining protocol accurately; inadequate fixation or improper staining technique can lead to poor visualization of bacteria
- Alcian yellow binds to the acidic components of the bacterial cell wall, while toluidine blue binds to the tissue background
- Use chemically clean glassware to avoid contamination
- Uses a light microscope
Periodic Acid-Schiff (PAS)
- This stains fungi and certain bacteria (e.g., Candida species, Tropheryma whipplei)
- Formalin is the most desirable fixative
- Ensure the tissue is adequately washed and processed to remove any residual fixative that may interfere with staining if another fixative has been used
- Periodic acid oxidizes carbohydrates to aldehydes
- Schiff reagent reacts with aldehydes to produce a magenta color
- Fungi and certain bacteria appear magenta against a light pink or colorless background after staining
- Tissue known to be infected with fungi or specific bacteria like Candida species or Tropheryma whipplei is appropriate control material
- Ensure proper oxidation and follow staining protocol accurately; inadequate oxidation or improper staining technique can lead to poor visualization
- Periodic acid oxidizes carbohydrates in the cell walls of fungi and bacteria, and Schiff reagent binds to the resulting aldehydes, producing a magenta color
- Use chemically clean glassware to avoid contamination
- Uses a light microscope
Chromic Acid-Schiff (CAS) Stain Details
- This stains fungi (e.g., Histoplasma species)
- 10% neutral buffered formalin is the most desirable fixative
- Ensure the tissue is adequately washed to remove any residual fixative that might interfere with the staining process if another fixative has been used
- Chromic acid oxidizes polysaccharides in fungal cell walls
- Schiff reagent reacts with aldehyde groups formed by oxidation to produce a magenta color
- Fungal elements (e.g., Histoplasma species) will appear magenta against a light pink or colorless background after staining
- Tissue known to contain fungal elements, such as a sample with confirmed Histoplasma infection is appropriate control material
- Incorrect timing and concentration of chromic acid can cause Incomplete oxidation
- Adjusting the time in Schiff reagent can fix overstaining or understaining
- Chromic acid oxidizes polysaccharides in fungal cell walls to aldehydes, which then react with Schiff reagent to produce a magenta color
- Use chemically clean glassware to avoid contamination that could interfere with the staining reaction
- Uses a light microscope
Gridley Fungus Stain Detail
- This stains fungi (e.g., Aspergillus species)
- 10% neutral buffered formalin is the most desirable fixative
- Ensure the tissue is adequately washed to remove any residual fixative that might interfere with the staining process if another fixative has been used
- Chromic acid oxidizes polysaccharides in fungal cell walls
- Schiff reagent reacts with aldehyde groups formed by oxidation to produce a magenta color
- Aldehyde fuchsin stains fungal elements
- Fungal elements (e.g., Aspergillus species) will appear deep purple to black against a yellow background after staining
- Tissue known to contain fungal elements, such as a sample with confirmed Aspergillus infection is appropriate control material
- Incorrect timing and concentration of chromic acid can cause incomplete oxidation
- Adjusting the time in Schiff reagent and aldehyde fuchsin can fix overstaining or understaining
- Chromic acid oxidizes polysaccharides in fungal cell walls to aldehydes, which then react with Schiff reagent and aldehyde fuchsin to produce a deep purple to black color
- Use chemically clean glassware to avoid contamination that could interfere with the staining reaction
- Uses a light microscope
Grocott Modification of Gomori Methenamine-Silver Stain Details
- This stains fungi (e.g., Pneumocystis jirovecii)
- 10% neutral buffered formalin is the most desirable fixative
- The tissue can be post-fixed in 10% neutral buffered formalin if another fixative has been used
- Methenamine-silver nitrate solution stains fungal cell walls black
- Chromic acid oxidizes polysaccharides in fungal cell walls
- Sodium bisulfite removes excess chromic acid
- Gold chloride tones the silver stain
- Sodium thiosulfate removes unreacted silver
- Fungal elements, including Pneumocystis jirovecii, appear black against a green or pale yellow background after staining
- Tissue known to contain fungal elements is appropriate control material
- Ensure proper timing and reagent preparation; over-staining or under-staining can occur
- Inadequate oxidation can result in weak staining
- The chromic acid oxidizes polysaccharides in the fungal cell walls, which then react with the methenamine-silver nitrate solution to form a black precipitate
- Use chemically clean glassware to avoid contamination and ensure accurate staining
- Uses a light microscope
Mayer Mucicarmine Stain Details
- This stains fungi and Cryptococcus species (bacteria)
- 10% neutral buffered formalin is the most desirable fixative
- The tissue can be post-fixed in 10% neutral buffered formalin if another fixative has been used
- Mucicarmine solution stains acidic mucopolysaccharides and capsule of Cryptococcus species red
- Weigert's iron hematoxylin stains nuclei black
- Metanil yellow counterstains background
- Acidic mucopolysaccharides and capsules of Cryptococcus species appear red, nuclei appear black, and the background appears yellow after staining
- Tissue known to contain Cryptococcus species is appropriate control material
- Ensure proper timing and reagent preparation: over-staining or under-staining can occur
- Inadequate differentiation can result in poor contrast
- The mucicarmine solution stains acidic mucopolysaccharides and the capsule of Cryptococcus species, while Weigert's iron hematoxylin stains nuclei and metanil yellow provides a contrasting background
- Use chemically clean glassware to avoid contamination and ensure accurate staining
- Uses a light microscope
Warthin-Starry Stain Details
- This stains spirochetes (e.g., Treponema pallidum)
- 10% neutral buffered formalin is the most desirable fixative
- Ensure tissue is well-fixed and consider postfixation in formalin if necessary if another fixative has been used
- Silver nitrate (impregnation) is used for staining spirochetes
- Hydroquinone (developer) develops the silver stain
- Spirochetes appear black against a pale yellow to light brown background after staining
- Tissue known to contain spirochetes is appropriate control material
- Control development time carefully; overdevelopment can cause non-specific staining
- Ensure proper washing steps to avoid background staining
- This is a silver impregnation technique where silver ions are reduced to visible metallic silver by the developer
- Use chemically clean glassware to avoid contamination
- Uses a light microscope
Dieterle Stain Details
- This stains spirochetes (e.g., Legionella species)
- 10% neutral buffered formalin is the most desirable fixative
- Ensure tissue is well-fixed and consider postfixation in formalin if necessary if another fixative has been used
- Silver nitrate (impregnation) is used for staining spirochetes and Legionella species
- Uranyl nitrate (sensitizer) sensitizes the tissue for better silver impregnation
- Hydroquinone (developer) develops the silver stain
- Spirochetes and Legionella species appear black against a yellow to brown background after staining
- Tissue known to contain Legionella or spirochetes is appropriate control material
- Control development time carefully; overdevelopment can cause non-specific staining
- Ensure proper washing steps to avoid background staining
- This is a silver impregnation technique where silver ions are reduced to visible metallic silver by the developer
- Use chemically clean glassware to avoid contamination
- Uses a light microscope
Steiner and Steiner Stain Details
- This stains Bacteria, including Spirochetes and Helicobacter pylori
- 10% neutral buffered formalin is the most desirable fixative
- Ensure tissue is well-fixed and consider postfixation in formalin if necessary if another fixative has been used
- Silver nitrate (impregnation) is used for staining bacteria, including spirochetes and Helicobacter pylori
- Uranyl nitrate (sensitizer) sensitizes the tissue for better silver impregnation
- Hydroquinone (developer) develops the silver stain
- Bacteria, including spirochetes and Helicobacter pylori, appear black against a yellow to brown background after staining
- Tissue known to contain the target bacteria is appropriate control material
- Control development time carefully; overdevelopment can cause non-specific staining
- Ensure proper washing steps to avoid background staining
- This is a silver impregnation technique where silver ions are reduced to visible metallic silver by the developer
- Use chemically clean glassware to avoid contamination
- Uses a light microscope
Pigments
- Endogenous pigments originate within the body
- Melanin (skin and hair color) and lipofuscin (age pigment) are examples
- Exogenous pigments come from outside the body
- Carbon from smoke/pollution (black lung deposits) and tattoo ink are examples
- Hematogenous pigments are derived from blood
- Hemoglobin (red pigment in red blood cells) and bilirubin (yellow pigment from red blood cell breakdown) are examples
- Anthracotic pigment is an exogenous pigment from inhaled carbon particles
- Pollution or smoking are typical sources, leading to black lung deposits
- Endogenous, nonhematogenous pigments are endogenous but not blood-derived
- Melanin and lipofuscin are examples
- Minerals are inorganic substances essential for bodily functions
- Calcium (bones/teeth) and iron (hemoglobin component) are examples
Argentaffin Reaction
- This reaction shows the ability of cells/tissues to reduce silver salts to metallic silver without an external reducing agent
- Mechanism: Substances like melanin or certain proteins directly reduce silver ions, forming a black or brown precipitate
- Argentaffin cells are found in the gastrointestinal tract
Argyrophil Reaction
- This reaction involves cells/tissues binding silver ions but needing an external reducing agent to precipitate metallic silver
- Mechanism: Argyrophil cells lack intrinsic reducing properties; after silver ion binding, an external reducing agent (like formalin) develops the silver into a visible precipitate
- Argyrophil cells are also found in the gastrointestinal tract and other tissues, but their staining requires an additional reduction step
Argentaffin vs Argyrophil Reactions
- Similarities: Both reactions use silver staining to visualize specific cells/tissues; they are used in histology to identify cells with particular chemical properties
- Differences: Argentaffin cells reduce silver ions on their own, while argyrophil cells require an external reducing agent for silver precipitation
Prussian Blue Stain Details
- Demonstrates ferric iron (Fe³⁺)
- Alcohol-based fixatives or neutral buffered formalin are the most desirable fixatives
- Washing the tissue thoroughly in water before staining can help if a non-alcoholic fixative has been used
- Potassium ferrocyanide and hydrochloric acid react with ferric iron to form an insoluble blue compound
- Ferric iron deposits appear as blue or blue-green granules after staining
- Tissue known to contain ferric iron deposits makes appropriate control material
- Ensure proper timing and reagent concentration to avoid overstaining/understaining
- Ferric iron and potassium ferrocyanide react, forming Prussian blue
- Use chemically clean glassware to avoid contamination
- Light microscope is used
Turnbull Blue Stain Details
- Demonstrates ferrous iron (Fe²⁺)
- Alcohol-based fixatives or neutral buffered formalin are the most desirable fixatives
- Washing the tissue thoroughly in water before staining can help if a non-alcoholic fixative has been used
- Potassium ferricyanide and hydrochloric acid reacts with ferrous iron to form an insoluble blue compound
- Ferrous iron deposits appear as blue or blue-green granules after staining
- Tissue known to contain ferrous iron deposits makes appropriate control material
- Ensure proper timing and reagent concentration to avoid overstaining/understaining
- Ferrous iron and potassium ferricyanide react, forming Turnbull blue
- Use chemically clean glassware to avoid contamination
- Light microscope is used
Schmorl Ferric-Ferricyanide Reduction Test Details
- Demonstrates reducing substances like melanin, lipofuscin, and argentaffin cells
- 10% neutral buffered formalin is the most desirable fixative
- Thorough washing is important to remove any residual fixative
- Ferric chloride and potassium ferricyanide are the primary reagents, reacting with reducing substances to form a blue or green color
- Positive staining results in a blue or green color
- Tissue known to contain melanin or argentaffin cells makes appropriate control material
- Inadequate washing of fixative can lead to non-specific staining; ensure thorough washing steps
- Ferric-ferricyanide reacts with reducing substances in the tissue, resulting in a color change
- Use chemically clean glassware to avoid contamination
- Light microscope is used
Fontana-Masson Stain Details
- Demonstrates melanin and argentaffin granules
- 10% neutral buffered formalin is the most desirable fixative
- Thorough washing is important to remove any residual fixative
- Silver nitrate binds to melanin and argentaffin granules, then reduced to visible metallic silver
- Positive staining results in black deposits
- Tissue known to contain melanin or argentaffin granules makes appropriate control material
- Control the exposure time carefully; overexposure to silver nitrate can lead to non-specific staining
- Silver nitrate binds to melanin and argentaffin granules and is reduced to metallic silver, which is visible under the microscope
- Use chemically clean glassware to avoid contamination
- Light microscope is used
Grimelius Stain Details
- Demonstrates argyrophil granules
- 10% neutral buffered formalin is the most desirable fixative
- Thorough washing is important to remove any residual fixative
- Silver nitrate binds to argyrophil granules, then reduced to visible metallic silver using an external reducing agent
- Positive staining results in black or dark brown granules
- Tissue known to contain argyrophil granules makes appropriate control material
- Control the exposure time carefully; overexposure to silver nitrate can lead to non-specific staining
- Silver nitrate binds to argyrophil granules and is reduced to metallic silver, which is visible under the microscope
- Use chemically clean glassware to avoid contamination
- Light microscope is used
Churukian-Schenk Stain Details
- Demonstrates argyrophil granules
- 10% neutral buffered formalin is the most desirable fixative
- Thorough washing is important to remove any residual fixative
- Silver nitrate binds to argyrophil granules, then reduced to visible metallic silver using an external reducing agent
- Positive staining results in black or dark brown granules
- Tissue known to contain argyrophil granules makes appropriate control material
- Control the exposure time carefully; overexposure to silver nitrate can lead to non-specific staining
- Silver nitrate binds to argyrophil granules and is reduced to metallic silver, which is visible under the microscope
- Use chemically clean glassware to avoid contamination
- Light microscope is used
Gomori Methenamine-Silver (GMS) Stain Details
- Demonstrates basement membranes and fungi
- 10% neutral buffered formalin is the most desirable fixative
- If another fixative has been used thorough washing is important to remove any residual fixative
- Methenamine-silver nitrate solution stains basement membranes and fungi black
- Chromic acid oxidizes tissue components
- Sodium bisulfite removes excess chromic acid
- Gold chloride tones the silver
- Sodium thiosulfate removes unreacted silver
- Basement membranes and fungi appear black, while background tissue appears light green
- Tissue known to contain basement membranes and fungi, such as kidney or lung tissue makes appropriate control material
- Ensure proper timing and concentration; over-oxidation with chromic acid can lead to non-specific staining
- Ensure thorough washing steps; incomplete washing can result in background staining
- Silver ions are reduced to metallic silver by aldehyde groups in the basement membranes and fungi, resulting in a black deposit
- Use chemically clean glassware to avoid contamination and ensure accurate results
- Light microscope is used
Hall (Fouchet) Stain Details
- Demonstrates bilirubin
- 10% neutral buffered formalin is the most desirable fixative
- If another fixative has been used thorough washing is important to remove any residual fixative
- Fouchet's reagent oxidizes bilirubin to biliverdin, which is green
- Van Gieson's stain counterstains collagen and muscle fibers
- Bilirubin appears green, collagen appears red, and muscle fibers appear yellow
- Liver tissue known to contain bilirubin makes appropriate control material
- Ensure proper timing and concentration; over-oxidation can lead to loss of bilirubin staining
- Ensure thorough washing steps; incomplete washing can result in background staining
- Fouchet's reagent oxidizes bilirubin to biliverdin, which is green
- Use chemically clean glassware to avoid contamination and ensure accurate results
- Light microscope is used
Von Kossa Stain Details
- Demonstrates calcium
- 10% neutral buffered formalin is the most desirable fixative
- Decalcify the tissue if necessary
- Silver nitrate solution reacts with calcium to form black deposits
- Sodium thiosulfate removes unreacted silver
- Nuclear fast red counterstains
- Calcium deposits appear black, nuclei and background appear red
- Tissue known to contain calcium deposits makes appropriate control material
- Insure proper fixation and decalcification; incomplete fixation or decalcification can cause errors
- Silver ions replace calcium ions, forming silver phosphate which is then reduced to metallic silver
- Use chemically clean glassware to avoid contamination
- Light microscope is used
Alizarin Red S Stain Details
- Demonstrates calcium
- 10% neutral buffered formalin is the most desirable fixative
- Decalcify the tissue if necessary
- Alizarin red S solution binds to calcium, forming an orange-red complex
- Calcium deposits appear orange-red after staining
- Tissue known to contain calcium deposits makes appropriate control material
- Insure proper fixation and decalcification; incomplete fixation or decalcification can cause errors
- Alizarin red S binds to calcium ions, forming a colored complex
- Use chemically clean glassware to avoid contamination
- Light microscope is used
Rhodanine Stain Details
- Demonstrates copper
- 10% neutral buffered formalin is the most desirable fixative
- Ensure tissue is properly fixed and processed
- Rhodanine solution binds to copper, forming a red complex
- Mayer's hematoxylin counterstain
- Copper deposits appear red, nuclei appear blue after staining
- Tissue known to contain copper deposits makes appropriate control material
- Incomplete fixation or processing can cause errors, ensure proper fixation and processing
- Rhodanine binds to copper ions, forming a colored complex
- Use chemically clean glassware to avoid contamination
- Light microscope is used
Prussian Blue vs Turnbull Blue
- Prussian Blue Reaction detects ferric iron (Fe³⁺)
- Ferric ions react with potassium ferrocyanide in acidic conditions, forming insoluble Prussian blue that appears as blue deposits
- Tissue known to contain ferric iron, like spleen or liver tissue with hemosiderin deposits can be used for control
- Turnbull Blue Reaction detects ferrous iron (Fe²⁺)
- Ferrous ions react with potassium ferricyanide in acidic conditions, forming insoluble Turnbull blue that appears as blue deposits
- Tissue known to contain ferrous iron can be used for control
- Key Differences
- Prussian blue detects ferric iron (Fe³⁺) and uses potassium ferrocyanide
- Turnbull blue detects ferrous iron (Fe²⁺) and uses potassium ferricyanide
Melanin Locations
- Skin: Present in the epidermis, giving skin its color
- Hair: Found in the hair follicles, determining hair color
- Eyes: Present in the iris, giving it color and providing protection against UV radiation
Granules and Minerals Not Demonstrated By the Above Techniques
- Calcium Oxalate
- Method: Pizzolato Method
- Primary Reagents: Pizzolato’s reagent (silver nitrate solution) which reacts with oxalate to form a black or brown precipitate
- Results: Calcium oxalate appears as black or brown deposits
- Copper (other than Rhodanine)
- Method: Rubeanic Acid Method
- Primary Reagents: Rubeanic acid (dithiooxamide) which binds to copper, forming a green or blue complex
- Results: Copper deposits appear green or blue
- Urate Crystals
- Method: De Galantha Method
- Primary Reagents: Silver nitrate and methenamine, which react with urate crystals to form a black precipitate
- Results: Urate crystals appear as black deposits
- Amyloid
- Method: Congo Red Stain
- Primary Reagents: Congo red dye which binds to amyloid, giving it a red color under light microscopy and apple-green birefringence under polarized light
- Results: Amyloid deposits appear red under light microscopy and apple-green under polarized light
Method for Bleaching Melanin Pigment
- Fixation: Ensure the tissue is properly fixed, typically with 10% neutral buffered formalin
- Pre-treatment: Deparaffinize and hydrate the tissue sections through graded alcohols to water
- Bleaching Reagent: Prepare a bleaching solution, commonly using hydrogen peroxide (H₂O₂) or potassium permanganate (KMnO₄) followed by oxalic acid
- Hydrogen Peroxide Method: Use 3-10% hydrogen peroxide in distilled water
- Potassium Permanganate Method: Use 0.25-0.5% potassium permanganate followed by 1% oxalic acid to remove the brown precipitate
- Application: Apply the bleaching solution to the tissue sections for an appropriate time, generally 10-30 minutes
- Rinse: Rinse the sections thoroughly with distilled water to remove excess bleaching reagent
- Staining: Proceed with the desired staining protocol to visualize other tissue components
- Dehydration and Mounting: Dehydrate the sections through graded alcohols, clear in xylene, and mount with a suitable medium
Differentiation of Anthracotic Pigment From Other Dark Brown to Black Pigments
- Anthracotic Pigment: Appears as fine black granules, typically found in the lungs of individuals exposed to polluted air or smokers
- Test: Anthracotic pigment is insoluble in strong acids and alkalis, and it does not bleach with potassium permanganate or hydrogen peroxide
- Formalin Pigment: Appears as dark brown to black granules formed by the reaction of formaldehyde with blood
- Test: Formalin pigment can be removed by treatment with alcoholic picric acid
- Melanin Pigment: Appears as dark brown to black granules, found in skin, hair, and certain organs
- Test: Melanin pigment can be bleached with hydrogen peroxide or potassium permanganate followed by oxalic acid.
- In summary:
- Anthracotic pigment is insoluble in acids and alkalis and does not bleach
- Formalin pigment can be removed with alcoholic picric acid
- Melanin pigment can be bleached with hydrogen peroxide or potassium permanganate
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