1DMT MT631 Unit 5 (Part 3) Specialized Areas of MedTech PDF

Specialized Areas of MedTech - 3 MT 631 | Mr. Jude Anthony C. Trinidad | S.Y 2024-2025 i. Sedimentation TOPIC OVERVIEW ii. Floatation A. CLINICAL...

Specialized Areas of MedTech - 3 MT 631 | Mr. Jude Anthony C. Trinidad | S.Y 2024-2025 i. Sedimentation TOPIC OVERVIEW ii. Floatation A. CLINICAL CHEMISTRY C. HISTOPATHOLOGY a. Analytical Technique in Clinical a. Process Chemistry i. Accessioning i. Spectrophotometry ii. Fixation ii. Chromatography iii. Dehydration iii. Electrophoresis iv. Clearing iv. Ion-Selective Electrode v. Infiltration or (ISE) Analysis Impregnation v. Enzyme-linked vi. Embedding Immunosorbent Assay vii. Blocking (ELISA) viii. Trimming or Facing vi. Mass Spectrometry (MS) ix. Sectioning b. Diabetes-Related Tests x. Staining i. Fasting Blood Glucose xi. Mounting (FBG) xii. Labeling ii. Oral Glucose Tolerance D. CYTOLOGY Test (OGTT) a. Pap’s Stain iii. Hemoglobin A1c (HbAc) E. CLINICAL MICROSCOPY c. Lipid Profile a. Urinalysis d. Liver Function Tests i. Dipstick Urinalysis e. Kidney Function Test ii. Basic (Routine) f. Thyroid Function Test Urinalysis g. Cardiac Biomarkers b. Automated Reagent Strip Readers h. Endocrine Function Test c. Semi-Automated i. Electrolyte Test d. Fully Automated j. Enzymes i. Liver Enzymes CLINICAL CHEMISTRY ii. Pancreatic Enzymes iii. Muscle Enzymes iv. Cardiac Enzymes k. Quality Control in Clinical Chemistry l. Automation in Clinical Chemistry m. Pneumatic Tube Systems B. PARASITOLOGY a. Terms b. Examination of Blood Basic chemical laboratory technique, i. Thick and Thin Blood chemical laboratory safety, electrolytes and Smear acid-base balance, proteins, carbohydrates, ii. Blood Concentration lipids, enzymes, metabolites, endocrine Technique function, and toxicology. c. Examination of Fecal Specimens Deals with the chemical components of d. Concentration Techniques biological samples  Analytical Techniques acids) based on size and charge Types: Spectrophotometry ★ Gel Electrophoresis: Used for protein and Measures the absorbance of light DNA analysis To determine concentration ★ Capillary Electrophoresis: Offers higher Types: resolution and faster analysis ★ UV-Vis Spectrophotometry: Measures absorbance in the ultraviolet and visible range. ★ Flame Photometry: Measures light emitted by excited atoms (used for sodium and potassium analysis). Ion-Selective Electrode (ISE) Analysis Measures the concentration of specific ions in a sample using selective membranes Common Ions Tested: ★ Sodium (Na⁺) ★ Potassium (K⁺) ★ Chloride (Cl⁻) Chromatography ★ Calcium (Ca²⁺) Separates components in a mixture Based on their affinity to a stationary phase and mobile phase. Types: ★ Gas Chromatography (GC): Analyzes volatile compounds ★ High-Performance Liquid Chromatography (HPLC): Used for non-volatile compounds Enzyme-linked Immunosorbent Assay (ELISA) Uses antibodies to detect the presence of specific proteins, hormones, or other biomarkers Types: ★ Direct ELISA: Antigen is directly detected ★ Indirect ELISA: Detects antibodies in the Electrophoresis sample Separates molecules (proteins, nucleic 2  Mass Spectrometry (MS) Diagnoses and monitor Diabetes mellitus Analyzes samples by ionizing chemical compounds and measuring their Fasting Blood Glucose (FBG) mass-to-charge ratios Measures blood sugar levels after fasting (8 Applications: hours), used to diagnose diabetes. ★ Toxicology, drug testing, metabolic ★ Sample: serum (w/o anticoagulant) profiling Advantages: Oral Glucose Tolerance Test (OGTT) ★ High sensitivity and specificity Assesses glucose handling after drinking a glucose solution. ★ Sample: serum (w/o anticoagulant) Used in pregnancy and requires fasting Consists of multiple extractions 0 hour Given 1 hour 2 hours 100 mL 1st of 2nd 3rd collection glucose collection collection With FBS solution Hemoglobin A1c (HbA1c) Reflects average blood glucose levels over the past 2-3 months, used for diabetes management. Diabetes-Related Tests ★ Sample: Red Blood Cells (w/o anti-coagulant) ★ Red cells are important ★ Anticoagulant: EDTA ★ No fasting Lipid Profile 3  Alanine Aminotransferase (ALT) Elevated in liver damage or inflammation. More specific to the liver Aspartate Aminotransferase Often elevated in liver or heart damage. Can indicate either heart or liver problem Alkaline Phosphatase Elevated in liver, bile duct, or bone disease. Fasting is 10-12 hours Bilirubin Total Cholesterol High levels indicate liver dysfunction or Measures the overall cholesterol level. hemolysis (breakdown of red blood cells). Metabolized in the liver High-Density Lipoprotein ★ Photosensitive analyte → sample is "Good" cholesterol, helps remove excess covered with carbon paper cholesterol from the bloodstream. ★ High protein, low fat Kidney Function Tests Low-Density Lipoprotein "Bad" cholesterol, contributes to plaque buildup in arteries. Very Low-Density Lipoprotein Triglyceride Type of fat in the blood, high levels indicate risk for heart disease. *if LDL, VLDL, and Triglycerides are high, there is a high risk of heart problem Creatinine Byproduct of muscle metabolism, high levels indicate impaired kidney function. Liver Function Tests Blood Urea Nitrogen (BUN) Measures the amount of nitrogen in the blood, which reflects kidney function. *Creatinine and Blood Urea Nitrogen (BUN) are normally removed by kidneys Electrolyte Levels Sodium, Potassium, Chloride, and No fasting Bicarbonate are closely related to kidney Liver Enzymes: health 4  ★ Low potassium levels = the kidney prevents the release of potassium Creatine Kinase-MB (CK-MB) Enzyme elevated during heart muscle injury. Thyroid Function Tests B-type Natriuretic Peptide (BNP) Indicator of heart failure severity. Endocrine Function Test Tests for the different endocrine organs Thyroid-Stimulating Hormone (TSH) Key hormone for diagnosing hypothyroidism or hyperthyroidism. ★ Hypothyroidism → High TSH ★ Hyperthyroidism → Low TSH *inversely proportional T3 (Triiodothyronine) and T4 (Thyroxine) Measures active thyroid hormones, essential for metabolism and growth regulation. Hypothyroidism ★ Low levels of thyroid hormones, High TSH Hyperthyroidism ★ High levels of thyroid hormones, Low TSH Cortisol Measures the stress hormone, useful in Cardiac Biomarkers diagnosing Cushing’s syndrome or adrenal STAT (shorter turnaround time) procedures insufficiency. Parathyroid Hormone (PTH) Regulates calcium levels, important in diagnosing parathyroid disorders. Insulin Tests for insulin production and resistance, key in diabetes diagnosis. Troponins (TnI and TnT) Lowers glucose levels in the blood Released during a heart attack, crucial for ★ The only considered hypoglycemic diagnosing acute myocardial infarction hormone (heart attack). Must be processed immediately once Electrolyte Function Test received as it is an indication of heart attack 5  Liver Enzymes Alanine Aminotransferase (ALT) ★ Elevated in liver damage, particularly from hepatitis or cirrhosis ★ Specific to the liver Aspartate Aminotransferase (AST) ★ Found in the liver and heart ★ Elevated in liver and muscle damage Alkaline Phosphatase (ALP) ★ Elevated in liver disease, bone disorders, Sodium and bile duct obstructions Critical for fluid balance and nerve function Gamma-glutamyl Transferase (GGT) ★ Elevated in liver disease, particularly due Potassium to alcohol or bile duct issues Important for heart and muscle function, ★ Most sensitive enzyme to alcohol intake imbalances can lead to arrhythmias. ★ Pseudo False increase of potassium → Pancreatic Enzymes because of hemolysis Amylase Hemolysis is a pre-analytical error ★ Elevation suggests acute pancreatitis or ★ Increased potassium levels → panic value other pancreatic diseases indicating that there is a heart problem Lipase ★ More specific to pancreatitis than Chloride amylase Helps maintain acid-base balance ★ Increases in cases of acute and other chronic pancreatitis Bicarbonate Regulates pH in the blood Muscle Enzymes Creatine Kinase (CK) ★ Elevated in muscle damage, including Enzymes conditions like muscular dystrophy and rhabdomyolysis Rhabdomyolysis - broken muscle cells which releases liver enzymes and myoglobin Myoglobin is toxic to the kidney which leads to damage Aldolase ★ Measures muscle damage, particularly in conditions like myopathies Proteins that catalyze biochemical reactions in the body Cardiac Enzymes Purpose of Enzyme Tests: Creatine Kinase-MB (CK-MB) ★ Diagnose diseases affecting specific ★ Marker for heart muscle damage, used in organs (e.g., heart, liver, pancreas). diagnosing heart attacks ★ Monitor disease progression and Troponin treatment effectiveness ★ Highly specific for cardiac injury, a key End with “ase” marker for myocardial infarction (heart 6  attack) Lactate Dehydrogenase (LDH) ★ Increases in heart, lung, or liver tissue damage ★ A non-specific enzyme Non-specific Enzymes High levels of non-specific enzymes indicates damage to di erent organs Quality Control in Clinical Chemistry Importance: ★ Ensures the reliability and accuracy of results Types: ★ Internal Quality Control: Daily testing of PARASITOLOGY known standards done inside the Study of parasites laboratory Branch of biology concerned with ★ External Quality Control: Comparison with phenomenon of dependence of one living other labs, also known as Proficiency Test organism on another. Automation in Clinical Chemistry Benefits: ★ Increased speed and accuracy ★ Reduces human error ★ Can process large volumes of samples Instruments: ★ Chemistry analyzers (e.g., Beckman Coulter, Siemens) Integrated systems for simultaneous testing ★ Hookworm (Ancylostoma duodenale) Old World Hookworm (Vampires: feed on blood) Parasites Organisms that live on and obtain their nutrients from another organism Host Pneumatic Tube Systems The organism from which a parasite obtains Transfer of specimen its nutrition and/or shelter Quick and safe way to transport samples from one place to another much faster than Examination of Blood manually delivering a sample by a 1. Thick and Thin Blood Smear healthcare provider a. Drop of blood b. Common way 7  2. Blood Concentration Technique a. More sample Examination of Blood b. mL of blood c. More sensitive Examination of Fecal Specimens 1. Wet mounts - most common technique Thick and Thin Blood Smear/Film ○ Direct Fecal Smear Commonly used in the diagnosis of Most common Plasmodium spp. 2 Reagents: ★ The causative agent of malaria NSS (assesses parasite Thin Smear motility) and Iodine ★ Species identifications (Specificity) (Temporary Stain that kills ★ Spread over the slide in a thin layer, the parasite) yielding intact, non-overlapping cellular 2. Concentration Techniques - more elements sensitive ★ Uneven distribution 3. Permanently Stained Smears - can be ★ Plasmodium spp. is the causative agent kept permanently of malaria ○ Wheatley’s Trichrome Stain Thick Smear ○ Iron Hematoxylin Stain ★ Sensitivity (more blood) ○ Modified Acid-Fast Stains ★ Preferred for Diagnosis (Coccidian) ★ Concentrated in a small area that is many 4. Culture cell layers deep 5. Immunoassay (Antigen Detection) ○ Giardia spp. Concentration Techniques Based on: ★ Sedimentation ★ Floatation Blood Concentration Technique Knott’s Concentration Sedimentation ★ Blood + 2% Formalin = Centrifuge The heavier parasites settle to the bottom as ★ Sediment: Check for Microfilaria (young a result of the gravity or centrifugation. form of filaria worms) Example: Formalin Ether Concentration Technique 8  Floatation 10. Staining Lighter parasite cysts and eggs rise to the 11. Mounting surface of a solution of high specific gravity 12. Labeling Brine Flotation Accessioning Process of assigning an identification code for the specimen to properly distinguish it from others without the use of patient’s name ★ Example: 2024-002 Fixation Process of chemically preserving tissues and HISTOPATHOLOGY their cellular constituents in a condition that is Study of abnormal tissues somewhat similar to that occurring during life ★ Histology (the study of normal tissues) is The sample will rot if not preserved the opposite of histopathology ★ Common fixative: Formalin Derived from three Greek words: ★ Histos - tissues ★ Pathos - disease of suffering ★ Logos - study The study and examination of abnormal tissues in aid of identification and diagnosis Pathologists will select the part of the organ → the MT will process the organ → the Pathologist will examine Dehydration Process of removing excess water from the tissue using dehydrating agent, such as alcohol, prior to replacement by an impregnation medium, such as paraffin wax ★ This is done because water does not mix with wax Histopathology Process 1. Accessioning Clearing or Dealcoholization 2. Fixation Process of completely removing the 3. Dehydration dehydrating agent (alcohol) in the tissue and 4. Clearing or Dealcoholization replacing it with a fluid that is soluble to the 5. Infiltration or Impregnation paraffin wax that will used for tissue 6. Embedding impregnation 7. Blocking ★ Common reagent: Xylene 8. Trimming or Facing 9. Sectioning or Microtomy 9  exposed Done to concentrate the cutting to one portion Sectioning or Microtomy Infiltration or Impregnation Process of cutting the tissue blocks into micro-thin slices of tissue ribbons using a microtome Staining Process of premeating tissues with a solution or medium, such as paraffin wax, that will fill the natural spaces, holes/cavities, and interstices to provide rigid support for the tissues Embedding Process of coloring cellular components of the tissue with a variety of natural or synthetic dyes for the purpose of optically differentiating the various cellular and tissue constituents Mounting Process of forming a tissue block using a mold as a shaping device Blocking An optional process and is performed only when embedding is performed in certain types of embedding molds, such as the Compound Embedding Unit Process of securing a coverslip on top of a tissue slide section to protect the tissue Trimming or Facing section and facilitate the ease of handling Process of cutting or removing excess wax and storage of the slide from the tissue block. Done to avoid destroying the tissue Usually, a four-sided prism or truncated pyramid—opposite sides being parallel—is Labeling formed after trimming to fit the microtome block holder, and the tissue is partially 10  Performed in screening laboratories. Uses a dipstick or a reagent strip Basic (Routine) Urinalysis Adds a microscopic examination of urine sediment to the reagent strip urinalysis. Specimen Evaluations ★ Specimen acceptability Process of providing permanent specimen Gross/ Physical Examination identification with printed or electronic ★ Color scanned labels of the prepared glass slides for proper identification, indicating the specimen code/number is similar to the code used in accessioning CYTOLOGY Study of cells Papanicoulaou/Pap’s Stain Light yellow and yellow - normal Part of exfoliative cytology Dark yellow - dehydrated Study normal and abnormal cells for the Red - most common abnormal color evaluation of various infections, abnormal In men: can indicate presence of hormonal activities, and even precancerous kidney stones lesions. In women: indicates menstruation Performed to assess the vaginal area ★ Clarity Usually performed in older women or those who experienced pregnancy ★ Odor Fruity odor can indicate diabetes ★ Specific gravity ★ pH pH and specific gravity are read CLINICAL MICROSCOPY using a reagent strip/dipstick but Study of urine and other body fluids with the are not part of the chemical aid of microscope and other laboratory component of urine instruments. Chemical Screening Urinalysis ★ Reagent strip/dipstick Oldest laboratory procedure Results are based on colors requiring ★ Dipstick Urinalysis the MT not to color blind ★ Basic (Routine) Urinalysis Dipstick Urinalysis 11  Examination of Urine Sediments ★ Microscope ★ Detects the formed elements of urine ★ Squamous Epithelial Cell is the largest cell found in the urine which can indicate if a microscope is out or in focus. Automated Reagent Strip Readers Uses spectrophotometric measurement of light reflection. Uses the principle that light reflection from the test pads decreases in proportion to the intensity of color produced by the concentration of the test substance. Semi-Automated Still dependent on an operator for sample mixing, test strip dipping, and inputting of physical and microscopic results. Requires a Medical Technologist Fully Automated Add urine to the rgt strip, and automated urine cell analyzers mix, aspirate, dilute and stain urine to classify urine sediment particles. Examples: USTH Laboratory and MMC Laboratory 12