Principles of Medical Laboratory Science 1 PDF

Tab 1  Principles of Medical Laboratory Science 1 1 Lesson 2.1: Hematology Section Pluripotent ↳ A...

Tab 1  Principles of Medical Laboratory Science 1 1 Lesson 2.1: Hematology Section Pluripotent ↳ Ability of the stem cell to differentiate to other Hematology stem cells. Study of blood, blood-forming tissues and related ○ Myeloid (RBCs, WBCs) disorders of blood. ○ Lymphoid (Lymphocytes) Related Disorders B, T, and NK Cells ↳ Anemia (RBC) ↳ Leukemia (WBC) Blast Cells ↳ Dengue/CFD (Platelets) First identifiable immature cell stages of what Blood is a specialized connective tissue that would eventually become RBCs, WBCs, and consists of formed elements (hemocytes) platelets. Has several stages before maturation. ↳ 38-48% of total blood volume ↳ Myeloblast ↳ Red blood cells (RBCs/Erythrocytes) ↳ Lymphoblast ↳ White blood cells (WBCs/Leukocytes) ↳ Erythroblast ↳ Platelets (Thrombocytes) ↳ Megakaryoblast Hematopoiesis/Blood Cell Formation Note: Only mature cells enter the circulating blood Origin or the process of blood cell formation and development. Factors that Stimulate Production of Blood Cells Primarily occurs in red bone marrow 1. Intrinsic Factor ↳ Bone marrows can be red and yellow Helps with B12 (Cobalamin) absorption in Children: Most bones the digestive tract Adult: Flat bones B12 is critical for red blood cell formation ↳ Vertebrae 2. Thrombopoietin ↳ Ribs From the liver ↳ Iliac crest Stimulates production of platelets ↳ Sternum 3. Granulocyte-Macrophage Colony-Stimulating factor (GM-CSF) All forms of hemocytes come from red bone Increases formation of leukocytes marrow and all parts of myeloid stem cells. Fats, soft 4. Interleukins tissues, and cartilages are formed in the yellow bone Influence the growth and activation of the marrow. lymphocytes 1, 2, and 3 Stem Cells 5. Erythropoietin (EPO) Precursors of all cells Stimulates the bone marrow to produce Bone Marrow more red blood cells when needed in the body ↳ Can be triggered by Hypoxia ○ When the oxygen level is low in the Ability of Stem Cells tissues, the kidneys produce more Self Renewal erythropoietin to stimulate the ↳ Ability of stem cells to divide and produce differentiation and growth of erythroid more stem cells with the same properties. (erythrocyte)  2 6. Cytokines ↳ When too much retics, blood cells are Stimulates the production of leukocytes impaired Stem cells are the origin of all cells. In the bone Reticulum marrow, they are pluripotent and self renewal. Remnant RNA visualized as fine, thread-like strands when stained Types of Blood Cells in the Circulating Blood 1. Erythrocytes (Red Blood Cells) Reticulocytes Most numerous formed element in the Are released during: blood. 4.2-6.5 Million/mL. Measures 6 to 8 ↳ Hemorrhage microns. Primary function is to carry oxygen ↳ Massive blood cell destruction to all tissues. Contains hemoglobin 2. Leukocytes (White Blood Cells) ↳ Contains iron for oxygen transport and Play an essential role in immune response of gives reddish color to the blood the body ↳ Composed of: Are colorless and have nuclei ○ Iron Measure from 7 to 20 microns ○ Globulin (A type of Protein) 4,300 to 10,800 per cubic mL Arteries: Brighter Color, Venous: Darker Red Differentiated into two: RBCs do not have nuclei and are biconcave A. Granular WBCs in appearance ↳ Neutrophils (Lilac Pink), eosinophils ↳ Center is thinner than outer edge (Red Orange), basophils (Blue 120 day lifespan Black Granules) [BEN] ↳ RBC will undergo apoptosis B. Agranular WBCs (programmed cell death) that occurs in ↳ Lymphocytes (P and T Cells), a regulated and controlled manner to monocytes (Largest WBCs and eliminate unnecessary damage to the once it moves into tissue, it body. simulates into macrophage) ↳ After that, the RBCs will go to the spleen and others will be eaten by Granular WBCs WBCs as the start of bilirubin Granules contain digestive enzymes designed to metabolism. destroy or damage foreign cells Lobulated Reticulocytes/Retics ↳ Appearance of nucleus Immature RBCs ↳ Nuclei arranged in lobes or sections Exhibit a blue network when stained with a Polychromatic stain special stain ↳ Used to stain the nucleus and granules of Have been recently released from the bone WBCs marrow during hemorrhage and blood cell ↳ “Many colors” destruction and 1 to 2 days before it matures. ↳ Example: Wright’s stain Normal amounts are 0.5 to 1.0% Blue network is remnant RNA (Ribonucleic Acid) left in the cell when the nucleus expelled near the 1. Neutrophils end of the red blood cell maturation Measures 9-15 microns. Contains 3-5 lobes/. Gray/Pale purple granules  3 Also called as “Polymorphonuclear Measures 12-20 microns. Largest WBC. Leukocyte” or PMN due to multiple lobes. With “ground glass” and a pale blue or light Also called as “Seg” gray cytoplasm. First responders to bacterial infection Main function: To remove dead old cells and Phagocytosis has a 8-24 hour lifespan ↳ Primary function of neutrophils Diapedesis ↳ Act of engulfing and destroying foreign ↳ Process wherein monocytes travel invaders through vessel walls and out of the 10-24 hours lifespan circulatory system into the tissues Is a substance known as “Pus” When monocytes go to tissues, they are ↳ Dead WBCs called macrophages. Band Cell ↳ Immature form of neutrophil 2. Lymphocytes ↳ Appear in circulating blood during high Smallest. Contains dark, dense nuclei. demand for neutrophils Measure 7-19 microns. ↳ Increased during acute infection Later on mature into: ↳ Nucleus is C-shaped or S-shaped ↳ B cells - become plasma cells 2. Eosinophils ↳ T cells and Natural Killer cells - active Measures 10-16 microns. Bilobed nuclei (two in cell-mediated immunity lobes). With characteristic large orange/red 2nd predominant WBC in adult, usually granules. Detoxifies foreign substances and predominant in children has a 6 day lifespan. If high in adult = viral infection Increased in: Largest are atypical lymphocytes ↳ Allergic reactions ↳ Myocardial disease 3. Thrombocytes (Platelets) ↳ Parasitic infections Spherical or ovoid cell fragments. Measures 3. Basophils 0.5-3 microns. 150,000-450,000/mL. ⅔ of Measures 10-14 microns. Bilobed with large total platelets are circulating in the blood. ⅓ dark blue granules in the cytoplasm. Has a 6 are stored in the spleen day lifespan. Megakaryocyte Granules contain: ↳ Cytoplasmic fragments of large bone ↳ Heparin = Helps prevent blood from marrow cells clotting ↳ 8-10 day lifespan ↳ Histamine = Makes blood vessel walls more permeable for easier Hemostasis = Stoppage of bleeding phagocytosis ↳ Platelets stick together and adhere to Increased in the sides of wound to form a plug that ↳ Chronic infections, Healing of fills the opening infection, Allergic asthma, Contact Serotonin allergies ↳ Chemical that helps constrict blood vessels (vasoconstriction) Agranular WBCs Thrombus 1. Monocytes ↳ Blood clot within blood vessels when there is no vessel wall damage  4 ↳ Stops the flow of blood 5. Synovial Fluid ↳ Body tissues do not receive oxygen and 6. Seminal Fluid cells eventually die 7. Amniotic Fluid ↳ May cause heart attack (myocardial 8. Peritoneal Fluid infarction) or stroke Analysis of the Formed Elements Complete Blood Count (CBC) Most commonly ordered hematology test and is a comprehensive test providing information about all formed elements in the circulating blood Hematocrit Percentage of RBCs in a given volume of blood. Is the volume of RBCs packed by centrifugation in a given volume of blood Erythrocyte Sedimentation Rate Screening test for inflammation that measure the speed at which RBCs settle out of anticoagulated blood Lesson 2.2: Clinical Chemistry Clinical Chemistry Testing of analytes dissolved in the fluids A. Quantitative ↳ Provides actual number that represents the amount of substances present B. Qualitative ↳ Indicates presence or absence of specific chemicals Plasma Purpose of Testing Collected from a tube containing anticoagulant 1. To evaluate chemical changes Liquid portion of the blood in which blood cells 2. To diagnose disease and prescribe treatment are suspended 3. To monitor drug levels Makes up more than 50% of total blood volume 4. To identify the presence of substances Made up of 90% water containing dissolved substances Specimen Types Used for Analysis 1. Plasma 2. Serum Chemical compounds are: 3. Urine 1. Plasma proteins 4. Cerebrospinal Fluid  5 ↳ Remains in the blood as antibodies (immunoglobulins), blood-clotting proteins Clinical Chemistry Tests (fibrinogen and prothrombin) and albumin 1. Glucose ○ In the body, we have 5 ↳ Most commonly performed. To screen immunoglobulins; IgM, IgG, IgA, IgE, Diabetes Mellitus. To monitor glucose levels. IgD To diagnose Gestational Diabetes 2. Medications (Diagnosed during pregnancy). 3. Electrolytes ↳ Hb A1c (Glycosylated Hemoglobin) ↳ Compounds that have a positive or negative ○ Used for glucose monitoring. Indication charge when dissolved in water of overall blood sugar levels for the 4. Hormones past 3 months. Specimen is Whole ↳ From endocrine glands (pituitary and thyroid blood (collected in a lavender top tube gland) containing EDTA) 5. Lipids ○ No fasting requirement. ↳ Fat molecules (cholesterol) ↳ Fasting Blood Sugar (FBS) 6. Enzymes ○ Measures blood glucose after a person ↳ Protein-based molecules that catalyze has fasted for at least 6 to 8 hours. chemical reactions ○ Purpose is to screen for diabetes and 7. Food pre-diabetes. Normal range is 70 to ↳ Used for energy. Contains vitamins and 110 mg/dL. minerals. 2. Cholesterol and Lipid 8. Dissolved Gasses ↳ Lipids ↳ Carbon dioxide, oxygen, nitrogen ○ Substances including oils and fats. ○ CO2 is also present in the form of Insoluble in water. Soluble in organic bicarbonate for pH regulation solvents. 9. Waste ○ They are alternative sources of energy ↳ Processed in the liver and removed from the ↳ Cholesterol body through urinary system ○ Example of a lipid specifically a sterol and is a vital component of the cell Serum membrane. Liquid portion of blood collected from a tube ○ Used by the body to produce without anticoagulant. 90 to 92% water. hormones such as estrogen and Proteins (Albumin & Globulin) and other testosterone, Vit. D and Bile acids blood-clotting factors are utilized as the blood clot which aid in fat digestion. forms. Serum is commonly used because there is ○ Primarily serves structural and less protein interference, unlike plasma. regulatory roles in cells and is crucial for proper cellular function. Urine Quantitative and is used to screen for kidney damage caused by diabetes, hypertension, etc. Used to know the cause of renal calculi or kidney stone formation Specimen used: 24-hours urine specimen (timed specimen) ↳ Lipoproteins  6 ○ Protein molecules where lipid attaches C. Lipid Panels for transport ↳ Total cholesterol, Triglyceride, HDL, LDL, ↳ Types of Lipids: VLDL. Total cholesterol: HDL ratio. Provides ○ Cholesterol information for risk factors for coronary artery ○ Triglycerides - Correlated with heart disease diseases ○ HDL (High-density lipoprotein) - Good 3. Electrolytes ○ LDL (Low-density lipoprotein) - Bad ↳ Minerals in the body that carry an electric ○ VLDL (Very low-density lipoprotein) - charge (positive or negative) when dissolved Bad in water. A. Cholesterol ↳ Examples are: Sodium, potassium, chloride, ↳ Used for hormone production, absorption of magnesium, calcium, bicarbonate, and Vit. D, bile production and cellular membrane phosphate. structure. Also used as part of covering ↳ Provide information about: (myelin sheath) on some nerves of the body. ○ Acid-base balance of the body, Causes Fasting is not required because we have for edema, Status of kidney natural saturation for fats. disfunction, Dehydration ↳ Cholesterol: Non-fasting specimen while TG, HDL, LDL, VLDL: 12 hours fasting 4. Blood Urea Nitrogen (BUN) ↳ Endogenous cholesterol ↳ Measurement of urea in the blood. Its ○ Manufactured within the body primary role is to indicate waste that needs ↳ Exogenous cholesterol to be removed by the kidney. ○ Comes from diet. Found in animal ↳ Urea based food. Once consumed, dietary ○ Byproduct of protein metabolism, cholesterol is absorbed in the intestine Cleared by kidneys. and is packaged into lipoproteins, ○ If high = kidney disease specifically, chylomicrons. ○ If low = liver disease, low protein ○ Transports cholesterol through the intake, malnutrition bloodstream to various tissues or to the liver for processing. 5. Creatinine ↳ Plaque formation ↳ Byproduct of muscle metabolism, Correlated ○ Cholesterol build up with BUN and urine specimen. ↳ Atherosclerosis ↳ Creatine provides energy to muscle cells, ○ Plaque formation within blood vessels while creatinine is the byproduct of it. It is ○ Plaque may travel as an embolus filtered out by the kidney. causing blood clot ↳ High = indicates impaired kidney function ↳ Normal range for men 0.6 to 1.2 mg/dL for B. Triglycerides men and 0.5 to 1.1 mg/dL for women. ↳ Common in patients with a high 6. Thyroid Panels carbohydrate diet. Excess calories are ↳ Produced by thyroid gland for the regulation converted into triglycerides and stored as fat. of cellular metabolism ↳ Hyperlipidemia ↳ Thyroid hormones are: ○ High lipid (cholesterol and TG) ○ T3 (Triiodothyronine) ○ “Milky” serum/plasma > Lipemia ○ T4 (Thyroxine)  7 ↳ Thyrocalcitonin stimulates calcium storage ○ “Icteric” serum/plasma (dark ↳ TSH (Thyroid-stimulating hormone) yellow/greenish) ○ Produced by the pituitary gland ○ High liver enzymes ○ Stimulates T3 and T4 ↳ Tests: ○ High = hypothyroidism, Low = ○ ALT (Alanine aminotransferase) hyperthyroidism Non specific liver marker ↳ Iodine is essential for thyroid ○ ALP (Alkaline phosphatase) ↳ Hypothyroidism Indicates bile duct ○ Thyroid doesn’t produce enough obstruction and bone hormones to stimulate cell metabolism disease ↳ Hyperthyroidism ○ AST (Aspartate aminotransferase) ○ Overstimulation of cell metabolism Non specific liver marker 7. Cardiac Enzymes ○ Bilirubin ↳ Proteins are released into the bloodstream ○ Total Protein when the heart muscle is damaged. ○ Albumin ↳ Ordered in the lab during suspected heart ○ GGT (Gamma Glutamyl Transferase) damage and usually ordered in the ER. Helps diagnose bile duct and ↳ Ischemia liver issues and can indicate ○ Temporary decrease of blood flow to alcohol use disorders the heart muscle ↳ Myocardial Infarction Test Tube Color Fasting ○ Heart attack Requirement Cardiac Panel: a. Total Creatine Kinase (CK) HbA1c Lavender None ↳ Not only present in the heart FBS Gray 6-8 Hours b. Creatine Kinase Myocardial Band (CKMB) ↳ Isoenzyme specific for the heart Cholesterol Red / None ↳ Starts to rise 4 hours after MI, peak 18 hours Gel-Barrier / c. Troponin I Green ↳ Sensitive to myocardial damage LDL, VLDL, Red (Serum 12 Hours ↳ Elevates sooner than CKMB, lasts longer HDL, and TG Tube) ↳ Tested every 2-4 hours for the first 24 hours d. Myoglobin Electrolytes Green (Lithium None ↳ Found in cardiac and skeletal muscle Heparin Tube) ↳ Elevates sooner than Troponin ↳ Not specific BUN Green None / If other tests are done e. Brain Natriuretic Peptide (BNP) such as BMP ↳ Synthesized in ventricles of the heart or CMP, fasting ↳ To differentiate DYSPNEA from heart failure is 8-12 Hours 8. Hepatic/Liver Enzymes ↳ To monitor liver when patients are taking Creatinine Red / Creatinine medication causing damage to the liver Gel-Barrier / alone: None Green ↳ Jaundice Creatinine as part of a panel:  8 Clinical Microscopy 12 Hours Tests done are: Thyroid Panels Red / None ↳ Urinalysis (T3 and T4) Gel-Barrier / ↳ Sperm Analysis Green ↳ Analysis of Bodily Fluids CK Red / None Gel-Barrier / Urinalysis Green Fast, simple, inexpensive Reliable to detect or monitor the progress of CK-MB Serum None numerous diseases such as disorders of Separator / carbohydrate metabolism, UTI and abnormal pH Green / changes in the body Lavender Detects kidney and liver malfunction Troponin I Light Green / None Screening procedure for asymptomatic disease Green EBAM [Qualitative Reporting: Rare (0-1), few (2-3), occasional (5), moderate (5-10), many Myoglobin Plain Red None (more than 20) ] BNP Lavender / None ↳ Epithelial Cells Pink ↳ Bacteria ↳ Amorphous Urates ALT Red / ALT alone: ↳ Mucus Threads Gel-Barrier / None Green If part of a Parts of Urinalysis panel: 10-12 Physical examination, chemical testing, Hours microscope examination of urine sediments for detection of formed elements present. AST Red 10-12 Hours Quality Assurance for Urine Testing Procedures ALP Red / None Gel-Barrier / The first thing that must be checked is the Green integrity of the urine sample. Clean-catch midstream urine Bilirubin Green 4 Hours ↳ Best sample for urinalysis Sterile collection container (screw cap container) Total Protein Red / None Gel-Barrier / Analysis within 1 hour Green ↳ If not, refrigerate for 4-5 degrees Celsius Calibrate equipment by using QC Albumin Red / None Staff training Gel-Barrier / External QC Green Reagent strips must be stored properly with GGT Red / Green / 8 Hours desiccant and cap Light Green Types of Urine Lesson 2.3: Urinalysis/Clinical Microscopy Timed specimens  9 ↳ Post prandial specimens during glucose ↳ Presence of Myoglobin tolerance testing ○ Byproduct of muscle destruction 24-hour urine Blue or Green ↳ First urination is discarded, but the second ↳ UTI caused by Pseudomonas bacteria will be. Sample will be refrigerated. Fluorescence (Glowing) ↳ In depth analysis: creatinine, electrolytes, ↳ Multivitamins hormones. ↳ Normal Post Prostatic massage specimens Urine for Chlamydia testing Brown or Black Urine for drug testing ↳ Hemoglobin in acidic urine ↳ Presence of Melanin and Melanogen Physical Characteristics of Urine Urine Color Urine Clarity / Turbidity Urochrome Transparency ↳ Substance that provides yellow color Clear, slightly hazy, hazy, cloudy / slightly turbid, ↳ Product of Hemoglobin breakdown and turbid normal metabolism ↳ More cloudy = higher concentration of ↳ Higher concentration = Darker yellow formed elements Straw, pale yellow, dark yellow, amber Clear ↳ Straw = Colorless ↳ No visible particles, transparent ↳ You can report the color that signifies it ↳ Normal visually, or only the color itself. Hazy ↳ Few particles, not completely transparent Cloudy ↳ Difficult to see through ↳ Pyuria ○ Presence of WBCs ↳ Bacteria, mucus, RBCs, sperm, yeast or casts Dark Yellow to Amber Turbid ↳ Normal ↳ Signifies infection and inflammation. ↳ Presence of Bilirubin Urine Clarity ↳ Liver dysfunction ex: Hepatitis ↳ Refrigerated specimens may also become ↳ Increased RBC destruction cloudy. ↳ Strenuous exercise ↳ If urine becomes cloudy after refrigeration, it can be centrifuged. Orange ↳ Amorphous urates ↳ Bilirubin levels elevated ○ Acidic urine ↳ Due to Pyridium (Phenazopyridine) ○ Pink ○ Treatment for recurrent UTI ↳ Amorphous phosphates Red to Brown ○ Alkaline ↳ Hematuria ○ White, cloudy appearance ○ Presence of RBCs ↳ UTI or renal dysfunction  10 Specimen Volume ↳ Provides an approximate value for each Normal: 750-2,000 mL/day. 50 mL - preferred chemical specimen volume, 12 mL - minimum acceptable Reporting: volume ↳ Trace, 1+, 2+, 3+, or 4+ ↳ Test tube is 10 mL so the 2 mL is for grace Qualitative volume. ↳ Positive or negative Normal urination frequency is 6 to 8 times within 24 hours. Chemical Components of Urine Terms to describe urine volume abnormalities 1. Bilirubin ↳ Anuria = Severely reduced or complete ↳ Byproduct of RBC destruction. Early absence of urine output. It indicates the end indicator of liver disease or bile obstruction stage of renal disease, acute kidney injury, ↳ Created from Hemoglobin as part of dehydration, and blockage in the urinary degradation process tract. ○ Jaundice (Yellow-Orange Color) = High ↳ Urinary frequency bilirubin ↳ Oliguria = Low output of urine. Less than 400 ↳ It is synthesized in the liver and excreted in mL/day. the urine or feces. ↳ Polyuria = Excessive urine production: 2.5 to ↳ Is insoluble but it needs to be soluble to be 3 Liters of urine. excreted by the body. ↳ Enuresis = Involuntary urination ↳ Nocturia (Nocturnal Enuresis) = Night time 2. Blood urination a. Hematuria ↳ Urinary incontinence ↳ Presence of intact RBCs ↳ Cloudy, red, RBCs in urine sediment Urine Odor ↳ High RBC = Glomerulonephritis Ammonia odor b. Hemoglobinuria ↳ Due to urea degradation ↳ With Hemoglobin but without RBCs. RBC Bacterial contamination lysis in the urinary tract. Intravascular lysis Fruity or sweet odor (breakdown of RBCs within vessels) ↳ Presence of ketones ↳ Ex. Hemolytic transfusion reactions, Severe burns, Malaria, Hemolytic anemias Specific Gravity c. Myoglobinuria Density (weight) of urine compared to distilled ↳ Presence of myoglobin. Red-brown urine, water at the same temperature. The higher the clear. Kidney disease. dissolved substances, the higher the specific ↳ Ex. Muscle trauma, Convulsions, Alcoholism, gravity. Can be an early indication of renal Excessive exercise. dysfunction. Normal: 1.005-1.030 Distilled water: 1.000 Kidney can give up to 1.0040 3. Glucose ↳ High blood glucose 160-180 mg/dL. Chemical Characteristics of Urine Diabetes screening or renal failure. Fasting Semiquantitative urine or post-prandial urine. ↳ Glycosuria = Presence of sugar in urine  11 ↳ Glucosuria = Presence of glucose in urine ○ Elevated Bence Jones Protein ○ Coagulate: 40-60 C 4. Ketones ○ Dissolve: 100 C ↳ Product of fat metabolism ↳ Microalbuminuria ↳ Ketonuria ○ Chronic presence of small amount of ○ Ex: Diabetes Mellitus, Malabsorption, albumin Low carbohydrate intake, Vomiting ○ Albumin is the most common protein in ↳ 3 products released into the bloodstream as the urine fat is metabolized: ○ Ex: diabetics, strenuous exercise, ○ Acetone dehydration ○ Acetoacetic acid ○ Beta-hydroxybutyric acid 9. Urobilinogen ↳ To monitor patients with Type I DM ↳ Produced in the intestine by the intestinal bacteria as bilirubin is broken down 5. Leukocytes ↳ Liver is incapable of processing urobilinogen ↳ Indicative of inflammation or infection or in the blood urinary tract ↳ Suggests pyelonephritis 10. Specific Gravity / Density ↳ Leukocyte Esterase ↳ For monitoring hydration status ○ Substance present in granulocytic ↳ Pads: 1.000-1.030 WBCs detected by reagent strips ↳ Neutrophils ○ Common WBC in UTI 6. Nitrite ↳ Presence of bacteria ↳ First morning urine sample ↳ Escherichia coli ○ Most common cause of bacterial UTI ○ Others: Klebsiella, Proteus, Serratia ○ Specimen of choice: First monitoring specimen 7. pH ↳ “Parts Hydrogen” ↳ Normal: 4.6-8.0 Microscopic Examination of Urine ↳ Acidosis: blood pH is less than 7.35 Epithelial Cells ↳ Alkalosis: blood pH is greater than 7.45 Shed from lining of urinary tract 3 types 8. Protein ↳ Squamous epithelial cells (SEC) ↳ Proteinuria. Can make urine cloudy/turbid. ○ “fried egg: appearance ↳ Can be present even without renal disease ↳ Transitional Epithelial Cells (TEC) ↳ Multiple Myeloma ○ High in malignancy and catheterization ○ Cancer of bone marrow ↳ Renal Tubular Epithelial Cells (RTE)  12 ○ High in glomerulonephritis, viral infection, kidney disease Mucus Thread-like protein strands Not clinically significant Large amounts = positive protein result Blood Cells RBC ↳ Colorless, smooth, no nucleus Crystals WBC Formed from dissolved substance especially ↳ Granular salts ↳ Easier to identify Affected by pH, low temperature, high SG ↳ Pyuria ○ Presence of WBC in urine Spermatozoa May be present in urine of males and females Oval head (teardrop shape) and long tail If high in urine=positive for protein Important ↳ Retrograde Ejaculation ○ Sperm is released into the bladder instead of the urethra Artifacts Microorganisms Contaminants You report the genus and species, Fecal material, toilet paper, air bubble, scratch or number/quantity, stage of development. powder, clothing fibers Bacteria Often confused with other formed elements ↳ Usually result of external contamination ↳ May be in the form of bacilli or cocci Casts Fungi Formed due to reduced urine flow, increased ↳ Candida albicans amounts of dissolved substances and acidic ○ Most common fungi in urine urine conditions ○ “Candidiasis” Assume the shape of tubule or duct where they ○ Budding/branching appearance are formed Parasites May indicate blockage of the duct. ↳ Trichomonas vaginalis ○ STD ○ Vaginal inflammation for females, symptomatic for males. ○ Motile  13 Lesson 3.1: Immunology & Serology Immune Process: Three Defense Mechanisms Immunity [Innate] First Line of Defense: Mechanical & Recognition and destruction of foreign Chemical substances. Keep invaders out/prevent them from surviving long. Immunology Intact skin and mucous membranes. Study of components of the immune system and ↳ Physical and mechanical barrier to function. pathogens. ↳ Ex. adjusting pH, cilia, reflexes, acidic Serology stomach, enzyme in saliva, earwax, Area of the laboratory where antigen-antibody perspiration, tears (lysozyme) reactions are studied. Normal Flora Focuses on serum constituents. Because ↳ Competitive environment in which bacteria antigens and antibodies are found in the cannot survive. serum. [Innate] Second Line of Defense: Internal Antigen Nonspecific Response Any substance that triggers an immune Natural or innate immune response. response. Phagocytosis, inflammation, fever, and Natural Substances that have molecular weight below Killer (NK) cells. 100,000 Daltons. ↳ Histamine (vasodilators) is released when Immunogen = foreign substances greater than injured. It is a naturally occurring chemical 100,000 Daltons. compound that plays a role in the immune ↳ Able to produce adaptive specific immune response, released by mast cells during responses allergic reactions / response to injury. Phagocytosis Antibody (Ab) I. Chemotaxis Substances produced by the body in response to ○ Signal released by cells when injured antigens. ○ Activate WBC to act as phagocytes (eosinophils, dendritic cells) to undergo Globulin phagocytosis. Protein that makes up an antibody IV. Inflammation ○ Red, hot, swollen Immunoglobulin ○ Caused by pathogen infection Attachment and destruction of invader ○ Histamine Remembering identity Released by cells when irritated or 2 characteristics of adaptive immune response: injured memory and specificity “Leaky”, swelling, pain ↳ IgA = secretion ○ Pus ↳ IgG = chronic Cell debris (dead phagocytes, ↳ IgM = acute fluid, pathogen, injured blood cells) ↳ IgE = parasitic infection / hypersensitivity ↳ IgD = idiopathic  14 Third Line of Defense: Acquired/Adaptive Immunity Recognition - identify Memory - recognize in the next exposure ↳ Develops 1-2 weeks T Cells - Cell-mediated immunity B Cells - Antibody mediated immunity / humoral immunity V. Fever ○ Phagocytes signals brain to rise Cell-Mediated Immunity temperature Begins at the site of invasion ○ Lowers ability of pathogens to survive T cells become “Activated” when Ag is presented by macrophage 2 Groups of Protective Proteins 4 identical copies: Interferons ↳ Killer T Cells (CD8) - kill Ag ↳ Secreted by cells that have been infected by ↳ Helper T Cells (CD4) - stimulate T cells, viruses. helps B cells ↳ Resist spread and viral replication. ↳ Suppressor/Regulatory T Cells (CD8) - ↳ Are critical parts of the innate immune inhibit excess T and B cells system. ↳ Memory T Cells (CD4) - remembers ○ Type 1: alpha and beta. ○ Type 2: gamma. Destroys bacteria Antibody-Mediated Immunity through cell lysis. B cells Presented by macrophage Complement Proteins 2 Replicas: ↳ Destroy bacteria ↳ Plasma cells - secrete Abs, lock and key ↳ Holes in membrane ↳ Memory B cells - remember ↳ Jules Bordet discovered the complement proteins. 4 Ways to Acquire Immunity ○ According to activation: C1, 4, 2, 3, 5 Natural Active Immunity (membrane attack complex), 6, 7, 8, 9. From infection with a pathogen. Has previous ○ According to discovery: C1, 2, 3, 4, 6, exposure with a pathogen, therefore, the body 7, 8, 9 will produce antibodies on its own. Memory IV. Natural Killer (NK) Cells Long-term immunity ○ Special lymphocytes Ex. chicken pox ○ Attack and destroy infected human cells. They attack cells with improper Artificial Active Immunity cell division. From vaccines containing antigen to cause the ○ Ex. cancer cells body to develop antibodies. Cancer cells reside inside tissues Long-term so they are not recognized by our Ex. Hepatitis A and B, measles, mumps immune response.  15 Natural Passive Immunity 1939 on Rhesus monkeys From mother to infant through placenta or breast 85% population is Rh (+) milk. No encounter with antigens but the body gets its antibodies from another source. Erythroblastosis fetalis / Hemolytic Disease of the Newborn Temporary (HDN) Severe hemolytic anemia of newborns Artificial Passive Immunity Baby Rh (+), mother Rh (-) From injection of gamma globulins (artificial No effect on 1st pregnancy antibodies) from the mother, making the infant RhoGam inherit the antibodies. ↳ Anti-Rh immunoglobulin Short-acting ↳ Given during pregnancy and after delivery to For emergency situations Rh (-) mothers Failure of the Immune System Autoimmune Diseases Ex. rheumatoid arthritis Body can’t recognize self from non-self, therefore, the immune system kills its own body. A person can take immunosuppressant drugs to lower the immune response. Allergy Hypersensitive; life-threatening Inflammation and organ dysfunction Uses direct agglutination. Ex. insect stings, medications Lesson 3.2: Laboratory Safety Blood Types ABO Infection Control Blood Type RBC Plasma Percent in The process of protecting healthcare workers and Population patients from infectious agents A A Anti-B 40% Hazards, laws, safety equipment antigen Infectious agents are viruses, parasites, fungi, and bacteria. B B Anti-A 11% antigen Microorganisms O No Anti-A 45% Too small = microscope (universal antigen Anti-B Ex. bacteria, viruses, fungi, protozoa, and donor) parasites AB A and B No 4% Pathogens (universal antigen antibodies Disease-causing organisms recipient) Rh/D Antigen Infection Protein found on the surface of RBCs. Can be Invasion by pathogens that cause disease and positive or negative. symptoms  16 Fungi Contagious/Transmissible Plant-like Can be spread to other people/organisms Dark and damp Mycotic infections Types of Microorganisms ↳ Fungal infections often caused by yeast Bacteria ↳ Ex. athlete’s foot, ringworm Single-celled organisms with cell wall 4% are pathogens Parasites Antibiotics = medications that kill/keep bacteria Take nourishment from host from multiplying Larger multicellular organisms Neutrophils, monocytes (in circulation), and Ex. Malaria - attack RBCs, tapeworm, roundworm macrophages (in tissues) kill bacteria. When Killed by eosinophils. neutrophil cannot handle bacteria alone, monocytes/macrophage will come. Protozoa Under parasites Cocci Complex single-cell Round shaped Most are nonpathogenic Ex. staphylococci (grape-like clusters), Ex. amoeba streptococci (chains), diplococci (pairs). Sore throat, pneumonia, food poisoning Growth Requirements Sanitation Bacilli Reduces number of microorganisms Long, slender, rod-shaped Heat/chemical Spores ↳ Dormant form,resistant to changes in heat, Disinfection moisture and disinfectants Chemicals used to kill pathogens. Ex. botulism, tetanus, TB, Salmonella food Contact time must be 5 minutes. poison Skin disinfection: ↳ 70% isopropyl alcohol and Povidone-Iodine Spirilla Solution Curved/spiral-shaped Work surface/equipment: Less frequently isolated in the body ↳ 10% bleach solution Ex. Treponema pallidum ↳ Causative agent of syphilis Asepsis Without infection Viruses Smallest infectious agents Surgical Asepsis RNA/DNA with protein capsule All pathogens are destroyed before entering the Viewed by electron microscope body. Are killed by lymphocytes. Ex. venipuncture, injections, urinary Ex. measles, mumps, chicken pox, and HIV catheterization ↳ HIV = causative agent of AIDS, retrovirus Medical Asepsis Procedure/environment that allows a patient to be treated without exposure to pathogens.  17 Portal of Entry Elements for Microorganism Survival Mucous membranes 1. Temperature - body temperature 98.6 degrees External openings Fahrenheit / 37 degrees Celsius Same way as pathogens exit/leave the body 2. pH - neutral pH 3. Darkness and Moisture Susceptible Host 4. Nutrition - food source Not protected from pathogen as it enters the 5. Oxygen body of the host ↳ Aerobic: need oxygen to survive Age and illness are factors ↳ Anaerobic: absence or low oxygen to survive Precautions Chain of Infection Universal Precaution Infectious Agent Everyone is potentially infectious for bloodborne Pathogen: viruses, fungi, parasites, bacteria. pathogens and that same care should be used to Fomites = contaminated objects. treat every specimen. Yeast infection “Treat all specimens as potentially infectious” ↳ Developed from antibiotics overuse PPE Donning Sequence: lab gown, hair net, shoe ↳ Opportunistic pathogen cover, mask, goggles, gloves. PPE Doffing Sequence: gloves, handwashing, Reservoir Host removal of goggle, lab gown, hair net, shoe Infected/carrying the disease cover, mask Provides ideal environment for growth of pathogen Personal Protective Equipment (PPE) Gloves Portal of Exit Most important PPE in the medical laboratory The means by which the pathogen leaves the Properly-fitting body Wash hands before and after use Injection, ingestion, inhalation. Single use patient Through the eyes, mouth, ears, Prevent pathogen from entering the body intestinal/urinary/respiratory/reproductive tract, broken skin, blood or any other body fluids Mask To protect from droplets (coughing and sneezing) Mode of Transmission Should be kept tight Transfer of pathogens Through air, direct contact or through vectors Respirators (insects) For TB and other airborne diseases Vectors Gowns Living organisms that take in pathogens, allow it For isolation areas, nursery with newborns to live and multiply without becoming ill from it Interiors of gown should be free of contamination Dirty Hands Lab Coats Most common mode of transmission Use when there is a potential for splashing or soiling of clothes Fluid resistant  18 Note: Laboratory professionals should never apply ↳ QUality control makeup, eat, or drink when working with blood or ↳ Specimen processing other specimens. Point of Care Testing Food must never be stored in the same refrigerator Medical diagnostic testing performed outside the with specimens or medications, regardless of how it is clinical laboratory in close proximity to where the packaged. patient is receiving care. Often used by non-laboratory personnel Lesson 3.3: Current Issues in the Medical Devices are portable or hand-held Technology Profession Performed near the patient and immediate decisions can be achieved for the clinical Medical Engineering management of the patient Also called “Bioengineering” and “Biomedical ↳ Ex. glucometer - medical divide used to Engineering” determine approximate glucose Multi-disciplinary field integrating professional concentration engineering activities with a basic knowledge of ↳ Other examples: tests for hemoglobin, human body cholesterol, blood gases, and electrolytes Understanding how the body functions when it is health, diseased, or injured Genetic Engineering Also known as “Genetic Intervention” and Clinical Laboratory Instrumentation (Automation) “Genetic Reconstruction” Use of control systems and information Direct intervention in the genetic make-up of a technologies to reduce the need for human work living being, the replacement of genes or the in the production of goods and services addition of new genes to the genetic code or Can be applied to any or all of the steps used to blueprint of an organism. perform a manual laboratory assay Genetic Screening Benefits of Automation Medical procedure whose main purpose is to Reduction of medical errors detect genetic disorders through laboratory Improved safety for the laboratory professions screening Faster turn around time of results Usually carried out in Molecular Biology or Partially alleviating the impending shortage of Cytogenetics Laboratory skilled laboratory staff Detect presence of genetic disorder and chromosomal activities in a patient Laboratory Information System (LIS) Ex. Newborn screening - used to detect common Integration of computers through a common genetic disorders that can lead to mental database via various communication networks retardation or death if untreated Improves productivity Decreases risk for errors Prenatal Diagnosis Can perform: Employs a variety of techniques to determine the ↳ Data entry on patients’ charts health and condition of unborn fetus ↳ Data interpretation Congenital anomalies account for 20-25% of ↳ Inventory control perinatal deaths ↳ Online monitoring Helpful for  19 ↳ Managing remaining weeks of pregnancy ↳ Insemination - mixing sperm cell and egg ↳ Determining the outcome of pregnancy cell ↳ Planning for possible complication with birth Sperm cell fertilizes the egg cell a few hours after process insemination ↳ Planning for problems that may occur in the Intracytoplasmic Injection (ICSI) newborn infant ↳ Directly injecting the sperm cell into egg cell ↳ Deciding whether to continue the pregnancy if fertilization chance is low ↳ FInding conditions that may affect future pregnancies Step 4: Embryo Culture When fertilized egg divides, it becomes an In Vitro Fertilization (IVF) embryo Joining a woman’s egg cell and a man’s sperm Staff will regularly check if it is growing properly cell in a laboratory set-up Within 5 days, several cells are actively dividing In vitro means out of the body Fertilization connotes that sperm cell has Step 5: Embryo Transfer attached to and entered the egg cell 3-5 days after egg retrieval and fertilization Is a form of assisted reproductive technology Catheter containing the embryos will be inserted (ART) into vagina through the cervix, up into the womb ↳ Special medical techniques used to help a Pregnancy will result if the embryo is implanted in woman become pregnant the lining of the womb and grown Has been successfully performed and used since More than 1 embryo may be placed into the 1978 womb -> twins, triplets, etc. Used to treat cases of infertility such as: Steps in IVF ↳ Advanced maternal age Step 1: Stimulation of Superovulation ↳ Damaged or blocked fallopian tubes caused Fertility drugs will be given to woman to boost her by pelvic inflammatory disease or prior egg cell production reproductive surgery Normally, single egg cell per month ↳ Endometriosis (lining of uterus grows into Fertility drugs induce ovaries to produce several other places) eggs ↳ Male factor infertility, including decreased sperm count and blockage Step 2: Egg Retrieval ↳ Unexplained infertility Follicular Aspiration The Center for Advanced Reproductive Medicine ↳ Minor surgical procedure and Infertility (CARMI) of SLMC - Global City ↳ Performed to remove the egg cells from offers assisted reproductive technologies ovaries including IVF ↳ Thin needle is inserted through vagina into Sperm and Zygote Banking / Cryopreservation the ovary and sacs containing the eggs Sperm and zygotes are frozen and stored in ↳ Needle is connected to a suction device, liquid nitrogen cooled refrigerators pulls the eggs and fluid out, one at a time Sperm Banking = process of storing sperm in a sperm bank Step 3: Injection Therapeutic Sperm Banking = option available Sperm cell will be placed together with the best to man before starting radiation and quality egg cells and stored in an environmentally controlled chamber  20 chemotherapy who is concerned about possible Autografts side effect of infertility Organs or tissues that are transplanted within the Each sample is mixed with special media or same person’s body. solution to help provide protection during freezing and thawing Allografts Placed in plastic vials, coded, and frozen in Transplants that are performed between two liquid nitrogen vapor subjects of the same species Temperature of -196 C Can be either from a living or cadaveric source Zygote Banking Ex. heart, kidneys, liver, cornea, skin, etc. ↳ Process of storing zygotes in a zygote bank Most commonly transplanted organs are kidneys, ↳ Zygotes form after fertilization liver, and heart ↳ Purpose is to store zygote prior to transfer or Most commonly transplanted tissue is cornea placing to a woman’s womb Cloning 2 Types of Sperm and Zygote Banking Creation of an organism that is an exact genetic Short-Term Cryobanking copy of another Depositing, freezing and storage of sperm or Every single DNA is the similar zygote for less than a year First performed in 1997 for Dolly the sheep Recommended to preserve sperm or zygote for Natural cloning exists in the form of twins deferred inseminations Methods of Cloning Long-Term Cryobanking Artificial Embryo Twinning Performed to store sperm or zygote for more than Performed by separating a very early embryo into a year individual cells Allowing each cell to divide and developResulting Sex Prediction & Selection embryos are placed into a surrogate mother, Sex Prediction where they are carried and delivered Attempt to predict the gender of the offspring while it is till inside the mother’s womb Somatic Cell Nuclear Transfer (SCNT) Ultrasonography Performed by isolating a somatic cell from an organism and transferring the nucleus from that Sex Selection cell to a germ cell (egg or sperm) from which the Attempt of couples or individuals to control the nucleus had been removed. gender of offspring to achieve desired gender Pre-implantation methods are preferred and additional info : considered morally acceptable Color of top tubes Green - Osmotic fragility test Organ Transplantation Lavender - CBS Moving of an organ from one body to another or Yellow - Blood culture from a donor site on the patient’s own body, for Light blue - Coagulation studies the purpose of replacing the recipient’s damaged Black - ESR or absent organ. Red - FBS Cardinal signs of infection  21 Rubor -redness Plasma cells Dolor -pain Interleukins Calor -heat Immunoglobulins Tumor-swelling Antibodies Functio laesa -lost of function LAK cells Immunoglobulins Sections ? : IGA = secretion >Hematology: IGG = chronic * CBC IGM = acute * Hemoglobin and Hematocrit IGE = parasitic infection/ hypersensitivity (allergy) * Coagulation test IGD = idiopathic >Serology: * Antibody test Reagent strips * Hepatitis test Specific gravity - pka dissociation constants * HIV test PH - orensen's error of indicator >Clinical Microscopy: Glucose - double sequential enzyme reaction * Urinalysis Ketones - na nitroprusside reaction * Fecalysis Blood - pseudoperoxidase activity of hb * CSF and body fluid test Bilirubin - diazo reaction * Semen analysis Urobilinogen - Ehrlich's reaction: >Clinical Chemistry: Nitrite- Greiss reaction * Electrolytes test Leukocytes - Indoxyl carbonic acid ester + diazonium * Bilirubin test salt * Lipid profile >Blood Bank: Immuno responses * Blood typing >1st line of defense: * Pregnancy test HCl of the stomach Acidity of the vagina Immunity Peristalsis Active Natural: Infection Sneezing Active Artificial: Vaccination Skin Passive Natural: Transfer in vivo Lysozymes Passive Artificial: Immune serum Ig's administration >Second line of defense: WBC Description of urine NK cells Clear -no visible particles Inflammation Hazy -print blurred through urine Interferon gamma Cloudy -few particulates Antimicrobial substances Turbid -print cannot be seen through urine Milky Neutrophils -many precipitate or clot >Third line of defense: Chyluria -lymph fluid in urine CD4 CD8 Tests Suppressor cells Histopath -FNAB  22 Bacteriology -culture and sensitivity Clinical Chemistry -stone analysis Clinical Microscopy-body fluids Hematology -osmotic fragility testing Serology and Immunology -STD Panel Blood bank -crossmatching Reagent reading time Ascorbic Acid(ASC) -30s Glucose(GLU) -30s Bilirubin(BIL) -30s Ketone(KET) -40s Specific Gravity(SG) -45s Blood(BLO) -60s pH -60s Protein(PRO) -60s Urobilinogen (URO) -60s Nitrite(NIT) -60s Leukocytes(LEU) -120s