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6/26/2024 CEREBROSPINAL FLUID Chapter 10 PREAMBLE PowerPoints are a general overview and are provided to help students take notes over the video lecture ONLY. PowerPoints DO NOT cover the details needed for the Unit exam Each student is responsible for READING the...

6/26/2024 CEREBROSPINAL FLUID Chapter 10 PREAMBLE PowerPoints are a general overview and are provided to help students take notes over the video lecture ONLY. PowerPoints DO NOT cover the details needed for the Unit exam Each student is responsible for READING the TEXTBOOK for details to answer the UNIT OBJECTIVES Unit Objectives are your study guide (not this PowerPoint) Test questions cover the details of UNIT OBJECTIVES found only in your Textbook! 1  6/26/2024 C S F F O R M AT I O N AND PHYSIOLOGY Cerebrospinal fluid (CSF)  Supply nutrients to the nervous tissue  Removes metabolic waste Lining of the brain and spinal cord is made up meninges (3 layers)  Maintains intracranial pressure  Cushions the brain & spinal cord Dura mater (hard layer that lines the skull & spinal column) Arachnoid (spider web) filamentous inner membrane Pia mater – (gentle) thin lining membrane on the brain & spinal cord CSF is produced by the choroid plexuses (2 lumen ventricles and 3rd & 4th ventricles) capillary network of filtration between the blood plasma and CSF BLOOD-BRAIN BARRIER (tight fitting endothelial cells) 20 ml of CSF is produced every hour- fluid goes to the subarachnoid space Normal volume is 90 – 150 ml (neonates 10 – 60 ml C S F F O R M AT I O N AND PHYSIOLOGY The endothelial cells allow soluble nutrients and waste to exchanged between the plasma and tissues. The blood-brain barrier protects the brain. The valves act in a one-way response. Disruption of the barrier by disease (MENEGITIS) infection multiple sclerosis Allows protein, glucose and leukocytes (WBC) into the CSF 2  6/26/2024 SPECIMEN COLLECTION & HANDLING CSF collection = lumbar puncture between After the needle is in place, the opening pressure is recorded in 3rd & 4th vertebra or 5th & 6th vertebra patient chart by the trained physician performing the lumbar puncture. Elevated pressure requires the fluid to be removed slowly. Specimens are collected sterilely in 4 tubes 1. Chemistry and serology test – may be frozen 2. Microbiology – room temperature 3. Cell count – hematology – may refrigerate up to 4 hrs 4. Might be used for microbiology or for extra test CSF is STAT APPEARANCE OF CSF Report color and clarity Colorless / clear Cloudy Turbid ↑protein, lipids or WBCs Milky Xanthochromic = pink, orange, yellow = Due to RBCs breakdown, ↑ bilirubin Bloody (red) REFER to table 10-1 3  6/26/2024 BLOOD IN CSF Even or uneven distribution of blood Bloody = intracranial hemorrhage = even Uneven = traumatic tap highest concentration in tube 1 and diminish to tube 2, 3 If blood is present, there is a chance of clot formation. If a clot is present them fibrinogen (factor 1 ) is present. Fibrinogen should not be able to past the barrier therefore CSF should not clot Xanthochromic – is an indication of old blood (longer than a traumatic tap) Red = traumatic tap orange / yellow = old blood CELL COUNTS Cell counts leukocyte (WBC) or nucleated cells Normal adult values for CSF = 0 to 5 WBCs/ul children’s can be higher neonates = 30 mononuclear cells 200 wbc/ul and 400 rbc /ul - can still be clear Red blood cells Normal value for RBCs = 0, most red cells are due to the tap Automation is becoming more frequent for counting for standardization, precision and faster TAT Traditional way = Neubauer chamber and an equation Number of cells counted x dilution Number of squares counted x volume of 1 square = cells/ul 4  6/26/2024 CELL COUNTS Depending on the number of cells in the CSF W W Look on 10 x to see the distribution If few number of cells are present, count all 9 squares Sometimes, the WBC may be less than RBC and you may count in different areas W Stain can be used to help distinguish the WBCS Glacial acetic acid will lysis the RBCs to allow better visibility of the WBCs W DIFFERENTIAL COUNT After the total WBC is calculate, the cells need differentiated. Stained smear or cytospin slide The distribution of the cell should Cytocentrifuge be a monolayer. Forces cells onto a slide in a monolayer Filter paper absorbs moisture Some labs report only 0.1 mL CSF to 1 drop 30% albumin mononuclear and polynuclear Albumin increases the cell yield and decreases the cellular other report a 5-part differential distortion Table 10-2 gives recommendation to how many cells to count based off the chamber count 5  6/26/2024 DIFFERENTIAL COUNT Pleocytosis = increase of normal cells = ABNORMAL Normally Present – lymphocytes & monocytes Increased cells help diagnosis the meningitis ↑ neutrophils = Bacterial ↑ lymphocytes and monocytes = viral, tubercular, fungal or parasitic Refer to table 10-3 P O LY N U C L E A R C E L L S EOSINOPHIL NEUTROPHILS Parasitic infection Bacterial infections Fungal infection Early onset of viral, fungal, tubercular, parasitic Introduction of foreign material = allergic The appearance of a neutrophil reaction Cytoplasm vacuolated Granules could be lost may see phagocytized bacterial pyknotic = degenerated cells 6  6/26/2024 MONONUCLEAR CELLS LY M P H O C Y T E S MONOCYTES / MACROPHAGES Normal to see in low numbers Usually counted together Viral, tubercular, fungal meningitis Monocytes = blood Reactive lymphocytes Macrophages / histocytes = tissue Dark blue cytoplasm Viral, tubercular, fungal meningitis clumped chromatin plasma cell HIV infection (AIDS), multiple sclerosis, degenerative neurological disorders N O N PAT H O L O G I C A L LY A N D PAT H O L O G I C A L SIGNIFICANT IN CSF Lining cells = choroidal cells or Any for of a blast (1st stage of a hemopoietic cell) – ependymal cells seen in acute leukemias Lymphoma cells also can be seen Malignant cells – astrocytoma, retinoblastoma, medulloblastomas – usually dark, ugly and in clusters. Cells fuse together. 7  6/26/2024 CHEMISTRY TEST What tube is used? Tube 1 (Most common test Protein & Glucose) Filtrate of the plasma - Low molecular weight Reference ranges ≠ plasma values Protein = 15 – 45 mg/dL Albumin (predominately), prealbumin, α globulins =haptoglobin & ceruloplasmin β globulins = transferrin, carbohydrate deficient transferrin (TAU) Gamma globulins = IgG, IgA, IgM CHEMISTRY TEST Significance Decreased protein indicates a leak in CNS Elevated protein: Damage to blood-brain barrier Testing methodology = total protein 1. Turbidity (automated instruments)- nephelometry Immunoglobulin production in the CNS CSF/serum albumin index IgG index (compares the albumin to IgG) Neural tissue degeneration 2. Dye binding ability Oligoclonal bands (immunoglobins in CSF) See Box 10-1 Electrophoresis Myelin Basic Protein (from the myelin of the sheath surrounding the nerves) ↑ in trauma, encephalitis, Guillain-Barre, lupus and brain tumors 8  6/26/2024 CHEMISTRY TEST Glucose Selectively transported NORMAL RANGE is 60% - 70% of the plasma glucose CSF glucose should be compared to a serum glucose within 2hours of the tap 2 additional chemistry test (send out) Elevated glucose is related to a high serum glucose Lactate Decreased glucose = significant helps with diagnosis and managing meningitis ↓ glucose + ↑ WBCs (neutrophils) = bacterial >25 mg/dl = meningitis meningitis Hypoxia ↓ glucose + ↑ WBCs (lymphocytes) = tubercular Used to monitor bad head injuries meningitis Glutamine Normal glucose + ↑ WBCs (lymphocytes) = viral Produced by ammonia and α-ketoglutarate meningitis Normal 8 – 18 mg/dl Elevated in liver disease Table 10-4 MICROBIOLOGY TEST Aide in analyzing the CSF for organisms Micro test can take 24 hours to 6 weeks depending on the organisms CSF culture confirm results Preliminary test Grams stain & Acid-fast stain India ink Latex testing (on the way out) Molecular testing (nucleic acid amplification) PCR 9  6/26/2024 MICROBIOLOGY TEST Latex agglutination test / lateral flow assay – replaced India ink C. Neoformans Antigen panel – Strep. B, H. flu, Strep. pneumo., N. meng A, B, C, Y & W135, E coli K1 Naegleria fowleri – parasite in water source enters through the nose and migrates to the brain Serology test for syphilis = VDRL (similar to RPR in serum) P O S TA M B L E READ the TEXTBOOK for the details to answer the UNIT OBJECTIVES. USE THE UNIT OBJECTIVES AS A STUDY GUIDE All test questions come from detailed material found in the TEXTBOOK (Not this PowerPoint) and relate back to the Unit Objectives 10  6/26/2024 SEMEN Chapter 11 Preamble ◦ PowerPoints are a general overview and are provided to help students take notes over the video lecture ONLY. ◦ PowerPoints DO NOT cover the details needed for the Unit exam ◦ Each student is responsible for READING the TEXTBOOK for details to answer the UNIT OBJECTIVES ◦ Unit Objectives are your study guide (not this PowerPoint) ◦ Test questions cover the details of UNIT OBJECTIVES found only in your Textbook! 1  6/26/2024 ◦Semen consists of four components ◦ Contributed separately by the ◦ Testes and epididymis Physiology ◦ Spermatozoa produce in seminiferous tubules and mature in epididymis ◦ 5% of semen volume ◦ Seminal vessels ◦ Majority of fluid of semen, 60% ◦ Contains fructose – causes sperm motility ◦ Prostate ◦ Acidic fluid (acid phosphatase, citric acid) ◦ 20-30% of volume of semen ◦ Responsible for coagulation and liquefaction ◦ Bulbourethral glands ◦ 5% of volume of semen ◦ Alkaline fluid ◦ Normal semen specimen must have all four components Specimen Collection ◦ Reasons for testing – 1. fertility 2. to evaluate the success of vasectomy ◦ Proper collection is crucial for evaluation of fertility ◦First portion of the ejaculate is missing, then ◦ Sperm count will be decreased ◦ pH is falsely increased ◦ Specimen will not liquefy ◦Last portion of ejaculate is missing, then ◦ Semen volume is decreased ◦ Sperm count is falsely increased ◦ pH is falsely decreased ◦ Specimen will not clot 2  6/26/2024 Specimen Collection ◦ Requirements ◦ Abstinence at least for 2 days but not more than 7 days ◦ WHO – World Health Organization recommends 3 specimens between 7 days and 3 weeks (2 abnormal specimens = SIGNIFICANT) ◦ Detailed instructions should be given – written preferably ◦ Collection at the testing facility is preferred ◦ Delivery within 1 hour of collection at room temperature ◦ Positive identification ( name, birth date and date and time of collection) ◦ Collection should not have any nonspermicidial, condoms, or lubrication ◦ Should uses Standard Precautions Semen Analysis ◦ Macroscopic (TEST WITHIN 1 HOUR od collection) ◦ Appearance – Normal gray-white, translucent ◦ Abnormal = clear (sperm concentration low) = white (increased WBCs) = red (blood) = yellow (urine – which is toxic to sperm ◦ Liquification – fresh (clotted) should liquefy in 30 – 60 minutes. Clot >60 minutes = decreased prostatic enzymes ◦ Volume – Normal 2 – 5 mL – decreased volume could reflect improper collection ◦ Viscosity – consistent of fluid- draw up into pipette – report as 0 (watery) to 4 (gel-like) ◦ pH – Normal pH 7.2 – 8.0 - Increased pH = infection - Decreased pH = obstruction 3  6/26/2024 Semen Analysis ◦ Only one spermatozoon is needed to fertilize ◦ Various factors can affect the sperm ◦ Quantity – Normal >20 -250 M/mL, border-line 10 -20 M /mL ◦ Morphology of the head and tail ◦ Speed - Motility ◦ Microscopic ◦ Count on Neubauer chamber with a 1:20 dilution ◦ Diluting fluid is sodium bicarbonate & formalin ◦ Count in the four corners plus the center ◦ Side must match within 10% ◦ Automated instruments –Computer- assisted semen analysis ◦ Sperm Class analyzer ◦ CEROS CASA system ◦ Automated Sperm Quality Analyzer ◦ Motility Semen Analysis Microscopic continued ◦ Microscopic Motility – ◦ Need to have sperm with forward, progressive movement ◦ Well-mixed, liquefied semen specimen ◦ Examine within 1 hour; evaluate undiluted on glass slide with cover slip ◦ Estimate percentage with progressive, forward motion in 20 HP fields or, examine 200 sperm per slide and count the percentages of the different motile categories using a manual cell counter ◦ Grading can be done using a scale of 0 to 4, with 4 indicating rapid, straight-line movement and 0 indicating no movement ◦ A minimum motility of 50% with a rating of 2.0 after 1 hour is considered normal 4  6/26/2024 Semen Analysis Microscopic continued Additional Semen Analysis Sperm viability – mix with eosin-nigrosine stain to stain the dead sperm and count the number of dead per 100 sperm. Seminal fluid fructose energy for the sperm Normal ≥13 umol/ejaculate Anti-sperm antibodies – antibodies can be produced by male and female Microbial – Chlamydia, Mycoplasma & Ureaplasma Chemical – α-glucosidase, free L-carnitine, zinc, citric acid, glutamyl transpeptidase and prostatic acid phosphatase Post-vasectomy semen analysis Monthly testing beginning at 2 months Continues until no viable sperm are seen on a wet preparation Good Lab practice = Quality Control 5  6/26/2024 Postamble ◦ READ the TEXTBOOK for the details to answer the UNIT OBJECTIVES. ◦ USE THE UNIT OBJECTIVES AS A STUDY GUIDE ◦ All test questions come from detailed material found in the TEXTBOOK (Not this PowerPoint) and relate back to the Unit Objectives 6  6/26/2024 S Y N OV I AL F L U I D Chapter 12 PREAMBLE PowerPoints are a general overview and are provided to help students take notes over the video lecture ONLY. PowerPoints DO NOT cover the details needed for the Unit exam Each student is responsible for READING the TEXTBOOK for details to answer the UNIT OBJECTIVES Unit Objectives are your study guide (not this PowerPoint) Test questions cover the details of UNIT OBJECTIVES found only in your Textbook! 1  6/26/2024 PHYSIOLOGY Synovial fluid = joint fluid = viscous liquid of a movable joint. Knee, elbow, hip, shoulder Smooth articular cartilage & cavity with fluid Membrane lining = synoviocytes 2 types  Type A – macrophages-phagocytosis  Type B – fibroblast – produce hyaluronic acid, fibronectin and collagen An ultrafiltrate of plasma. Nonselective filtration – no large proteins, Reference ranges are like that of plasma- Table 12-1 Function Damage to membrane creates pain  Reduce friction and stiffness= arthritis  Lubricate the joint Common test are in Table 12-1  Nutrients to the cartilage  Reduces shock during compression during activities C L A S S I F I C AT I O N S Four classifications of disorders (arthritis) 1. Noninflammatory: degenerative, osteoarthritis 2. Inflammatory: immunologic, lupus erythematosus (LE), rheumatoid arthritis (RA), Lyme disease Crystal-induced, gout and pseudogout 3. Septic: microbial infection 4. Hemorrhagic: trauma, tumors, coagulation deficiencies 2  6/26/2024 COLLECTION AND HANDLING Needle aspiration = arthrocentesis Amount in joint is dependent on the size of the joint Normal knee 50% - Another chemistry test due to injury. A membrane disease would have a low hematocrit. Cholesterol serum to pleural Milky = chylous material (↑ triglycerides) >60 mg/dL = pseudochylous (↑ cholesterol) Serum / pleural ratio >0.3 Bilirubin serum to pleural Serum / pleural ratio >0.6 Glucose – helpful in rheumatoid arthritis Hematology Test Macrophages, neutrophils, lymphocytes, eosinophil, mesothelial, plasma cells, & malignant cells 64 – 80% macrophages Same in 18 – 30% lymphocytes pericardial & 1 – 2% neutrophils peritoneal 5  6/26/2024 ↑ neutrophils – infection, pancreatitis and pulmonary infarct. (b) Monocytes – macrophage, histocytes (a) Lymphocytes – small, large & reactive , increased in tuberculosis, viral infection, malignancy, autoimmune disease (c ) Eosinophil - >10% trauma to pleural cavity, allergic reactions & parasite Lining cells = Mesothelial. Can be pleomorphic. Reactive = clusters, varying amounts of cytoplasm, eccentric nuclei and multinucleated (may look like a malignant cells) reactive normal Malignant cells Large, ugly, dark, molding, no borders (cell walls) Irregular adenocarcinoma Small or oat cell carcinoma Metastatic breast cancer Only reported as suspicious cells refer to cytology Confirmed with special staining and flow cytometry 6  6/26/2024 Chemistry test on pleural fluid Most common test glucose, pH, total protein, adenosine deaminase, triglyceride & amylase ↓ glucose (,60 mg/dL) = tuberculosis, rheumatoid inflammation, malignant effusion, esophageal rupture pH 1000 = bacterial endocarditis 8  6/26/2024 Lab test Cell counts Chemistry – lactic dehydrogenase (LD) Markers levels for tumor Cultures – routine bacterial, fungal, acid-fast bacteria Peritoneal Fluid Accumulation of fluid in peritoneal membrane = ascites Calling it ascites fluid Sometimes after the procedure = paracentesis fluid Causes Hepatic disorders (cirrhosis) Bacterial infection in intestine (peritonitis) Peritoneal Lavage Diagnostic procedure to determine an intra-abdominal bleed Normal saline injected into cavity and withdrawn and cell count performed 9  6/26/2024 Peritoneal Fluid Transudate vs Exudate More difficult than other fluids Serum-ascites albumin gradient (SAAG) preferred over total protein and LD ratio Serum and fluid albumin levels are measured; fluid level is subtracted from serum level; difference (gradient) > than 1.1 is a transudate (hepatic origin) Serum albumin 3.8 – fluid albumin 1.2 = 2.6 = hepatic transudate Appearance Pale yellow / clear Turbid – bacterial infection Green / dark brown - bile Cellular Examination Normal nucleated cell 250 cell /uL or 50% of total WBC = Peritonitis or cirrhosis Cells: WBCs, mesothelial cells, macrophages (lipophages), malignant cells from various organs Yeast, Toxoplasmosis gondii 10  6/26/2024 Additional testing of Peritoneal Fluid Chemistry Microbiology & Serology Gram stains and aerobic and Glucose: below plasma levels = anaerobic cultures peritonitis and malignancy ↑ Amylase: pancrea s, Anaerobic cultures: inoculate gastrointestinal perforation blood culture bottle at bedside ↑ Alkaline phosphatase: Acid-fast smear, adenosine intestinal perforation deaminase and culture for TB ↑ BUN, crea nine: ruptured Tumor markers bladder, accidental perforation CEA CA 125 Presence of CA 125 antigen with a negative CEA suggests the source is from the ovaries, fallopian tubes, or endometrium Presence of CEA antigen suggests source is gastrointestinal Postamble READ the TEXTBOOK for the details to answer the UNIT OBJECTIVES. USE THE UNIT OBJECTIVES AS A STUDY GUIDE All test questions come from detailed material found in the TEXTBOOK (Not this PowerPoint) and relate back to the Unit Objectives 11  6/26/2024 B R O N C H OA LVE O L AR L AVAG E CHAPTER 14 A method for examining cellular, immunologic, and microbiological information from the lower respiratory tract Helpful for B R O N C H O A L V E O L A R L A V A G E ( B A L ) immunocompromised patients, patient with airway/ breathing problems 1  6/26/2024 SPECIMEN COLLECTION Procedure = Bronchoscopy – performed by Pulmonologist and the respiratory therapist Fiber optic into middle of lobe (upper or lower) Documentation of what area examined Sterile saline is slowly administered into the scope and mixed and then aspirated the contents for examination and culture The amount of saline instilled is 100 – 300 ml in 20 to 50 ml aliquots First sample in discarded An optimum sample is >30% of recovery (50-70%) 15 cells RBC difference should not be > 30 The cells should agree within 10% on each side Clumps present should be note “cell count may be inaccurate due to clumps of cells or clot” 3  6/26/2024 CELLS PRESENT IN BAL Evaluate the inflammatory process in the respiratory tract Cell that will be seen Macrophage Lymphocytes (CD4/CD8 ratio) Neutrophils Eosinophils Ciliated columnar bronchial epithelial cells Squamous epithelial cells CELLS PRESENT IN BAL Macrophage Lymphocyte Phagocytize Normal range is 1-5% Hemosiderin (golden, brown, black ↑ in lung disease, drug reactions, pigment within cell) pulmonary lymphoma & nonbacterial Foamy cells infection Normal range 56- 80% >25% lymphocytes - granulomatous lung disease >50% lymphocytes – hypersensitivity pneumonitis Ratio of CD4/CD8 will help define the process See Table 14-1 4  6/26/2024 CELLS PRESENT IN BAL Neutrophils Eosinophil Primary cell seen Usually, < 1-2% of total cells Normal is 25% eosinophilic lung disease ≥ 50% - acute lung injury, aspiration pneumonia or infection 5  6/26/2024 CELLS PRESENT IN BAL Erythrocytes (RBCs) Epithelial Cells Indicates acute alveolar hemorrhage or Ciliated columnar cells are prominent from procedure in bronch specimens Phagocytized RBC – suggest a alveolar Normal range in 4 – 17% hemorrhage in last 48 hours Hemosiderin macrophage indicates older than 48 hours M I C R O B I O LO GY TEST Box 14-1 list many different pathogens in the Bronchoalveolar Lavage Fungal, bacterial, and viral Quantification is helpful May be due to extended use of ventilator causing a pneumonia In the case with COVID PCR is useful in identification quickly 6  6/26/2024 CY TO LO GY Look for observation of : Sulfur granules (actinomyces) Hemosiderin-laden macrophage Langerhans cell Cytomegalic cells Fat droplets (Red O) Periodic-acid Schiff (PAS) for fungus Dust particles in pneumoconiosis or asbestos exposure PREAMBLE PowerPoints are a general overview and are provided to help students take notes over the video lecture ONLY. PowerPoints DO NOT cover the details needed for the Unit exam Each student is responsible for READING the TEXTBOOK for details to answer the UNIT OBJECTIVES Unit Objectives are your study guide (not this PowerPoint) Test questions cover the details of UNIT OBJECTIVES found only in your Textbook! 7  6/26/2024 P O STA M B L E READ the TEXTBOOK for the details to answer the UNIT OBJECTIVES. USE THE UNIT OBJECTIVES AS A STUDY GUIDE All test questions come from detailed material found in the TEXTBOOK (Not this PowerPoint) and relate back to the Unit Objectives 8  6/26/2024 Amniotic Fluid Chapter 15 Preamble PowerPoints are a general overview and are provided to help students take notes over the video lecture ONLY. PowerPoints DO NOT cover the details needed for the Unit exam Each student is responsible for READING the TEXTBOOK for details to answer the UNIT OBJECTIVES Unit Objectives are your study guide (not this PowerPoint) Test questions cover the details of UNIT OBJECTIVES found only in your Textbook! 1  6/26/2024 Physiology Amniotic fluid = cytogenetic Amniotic fluid is the product of the fetal metabolism – waste The fluid tells about the metabolism process & fetal maturation The fluid is in the amnion of the amniotic sac Function: Exchange of water and chemical between the fetus and the maternal circulation Cushion for the fetus Fetal movement Regulates temperature around the fetus Permits proper lung development Physiology continued Volume Regulates balance between production fetal urine, lung fluid, absorption of fetal swallowing and intramembranous flow (absorption of amniotic fluid water and solutes in the fetal vascular system) Fluid increase through the fetus development 12 weeks = 60 ml 3rd trimester = 800 – 1200 ml – secretes lung liquid in lung growth >1200 ml = polyhydramnios 2.0 mg/dL 3  6/26/2024 Maternal Urine versus Amniotic Fluid Needed to determine premature membrane rupture or accidental puncture of maternal bladder from amniocentesis Measure creatinine, glucose, protein, and urea Creatinine & urea will be lower in amniotic fluid than urine Amniotic fluid has 3000 mL) reaching large intestine = diarrhea Diarrhea Definition: >200 g stool weight per day with increased liquid and more than three movements per day Mechanisms of diarrhea: secretory, osmotic, altered motility Illness duration Water diarrhea has 2 types Mechanism’ 1. Secretory Severity 2. Osmotic types (fecal electrolytes) Stool characteristics Laboratory tests: fecal electrolytes, osmolarity, and pH Stool characteristics Normal fecal osmolarity = serum osmo >4-week duration = chronic Sodium, potassium, osmolarity used to calculate fecal osmotic gap 60 per hpf indicative of steatorrhea Split fat is more indicative 7  6/26/2024 Qualitative Fecal Fats NEUTRAL FAT SPLIT FAT 1. Homogenize one part stool 1. Mix specimen wit h ac et ic ac id a nd with two parts water. hea t 2. Mix emulsif ied stool with 2. Free fa tt y acids a nd fat t y acids from one drop 95% ethyl alcohol soa p hy droly sis an d n eu tral fats on slide. 3. Nu mber a nd siz e evaluat ed: normal up to 100 sma ll droplets( 60 per hpf indicative of corn ers steatorrhea Qualitative Fecal Fats Soaps and fatty acid do not stain on the split stain Neutral count the number of fat droplets Splits count the number and the size of the fat droplets Reported under high power f ield Normal =