Questions and Answers
Which mounting medium has the highest refractive index among the listed options?
What is the purpose of sealing the margins of the coverslip?
Which staining agent is known to act as a ripening agent and contains mercury?
Which antibody is commonly used in immunohistochemistry?
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What is the primary color of the nuclei after the acid alcohol differentiation step in the H & E staining process?
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What is the purpose of 0.5% acid alcohol in staining procedures?
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Which mordant is used in Weigert's hematoxylin staining?
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In the context of epithelial tumor markers, what does a positive CK 7 and negative CK 20 indicate?
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Which method is used to remove a coverslip from a section?
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What is the purpose of Eosin Y in histological staining?
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What does the abbreviation 'LUBO' in epithelial tumor markers refer to?
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Which staining method utilizes ammonia water as a blueing agent?
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What is the primary use of Acridine orange in histological staining?
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Which of the following fluids is specifically mentioned for use in frozen section preparations?
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Which type of slides do slotted staining dishes hold?
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What distinguishes Sudan III from Sudan IV in lysochrome staining?
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Study Notes
Stains and Reagents
- Ehrlich’s and Harris’ utilize iodate and HgO2 respectively as ripening agents.
- Iron hematoxylin uses iron as a mordant and is significant in staining processes.
Staining Techniques
- Weigert’s stain employs FeCl3 as a mordant, crucial for examining connective tissues and Entamoeba histolytica.
- Heidenhain’s stain incorporates ferric ammonium sulfate for mordanting.
- Mallory’s PTAH uses sunlight/K+ for staining fibrin.
- Copper hematoxylin specifically targets spermatogenesis.
Eosin Types
- Eosin Y (yellowish) is the most commonly used cytoplasmic/acidic stain.
- Eosin B (bluish) appears deep red, while ethyl eosin, Eosin S, and Eosin alcohol soluble variants offer flexibility in staining.
Staining Equipment
- Coplin jar holds 5-9 slides; slotted staining dishes accommodate 5-19 slides.
- Metal/glass staining racks can hold 10-30 slides for various staining procedures.
H & E Staining Steps
- Process begins with xylol for deparaffinization followed by a descending alcohol gradient for rehydration.
- Nucleus typically stains light blue, transitioning to blue with ammonia water, while red staining indicates cytoplasm.
Pap Smear Staining
- Hematoxylin serves as a nuclear stain; OG-6 and EA (Eosin Azure) are utilized for cytoplasmic staining of mature superficial and immature cells respectively.
Other Stains
- Benzidine indicates hemoglobin presence, acridine orange differentiates RNA and DNA via fluorescence.
- Congo red stains elastic tissue, amyloid, and myelin; malachite green detects Ascaris eggs.
Adhesives Used in Histology
- Mayer’s egg albumin, dried albumin, gelatin, and Poly-L-lysine are common adhesives, with 3-APES being noted for cytology.
Mounting Media
- Resinous media has a higher refractive index (e.g., DPX = 1.532), while aqueous media is typically used for lipid preservation.
- Glycerin jelly is the standard mounting medium for fat stains.
Slide Restaining and Maintenance
- Stain removal occurs with 0.5% acid alcohol and restaining techniques involve xylene and potassium permanganate.
- Broken slides can be salvaged by transferring sections to clean slides using a mixture of butyl acetate and durofix.
Ringing in Histology
- Sealing coverslip margins prevents fluid evaporation and immobilizes the coverslip while avoiding slide sticking.
Immunohistochemistry Insights
- Enzyme histochemistry primarily utilizes trypsin and protease; IgG is the most widely used antibody.
- Monoclonal antibodies are derived from mice, while polyclonal types originate from various animals, including rabbits and goats.
Epithelial Tumor Markers
- CK 7 is positive, and CK 20 is negative in tumors from lung, uterus, breast, and ovary (“LUBO” pattern).
- CK 20 positive and CK 7 negative characterize stomach and colon tumors; transitional cell carcinoma shows a dual positivity.
- EMA is positive in carcinomas, while CEA differentiates between adenocarcinoma and mesothelioma.
Additional Markers
- TTF-1 is a critical transcription factor for thyroid activity, relevant in examining associated carcinomas.
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Description
This quiz delves into various staining agents and mordants used in histology, including Ehrlich's, Gill's, and Delafield's. Learn the functions and applications of iron hematoxylin, eosin, and tungsten hematoxylin in staining techniques. Enhance your understanding of these essential reagents in histological preparations.
