EXAMINATION OF OTHER BODY FLUIDS MICROSCOPIC STRUCTURES

Elastic Fibers – Tuberculosis

Charcot-Leyden Crystals – Eosinophil degradation
SPUTUM – mucus secretion from the upper and lower
product  Bronchial Asthma
respiratory tracts.
Pigmented Cells – Contains carbon  Congestive Heart
o Mixture of plasma, electrolytes, mucin and
Failure
water.
o Heart-Failure Cells: Hemosiderin-laden
Macrophages
SPECIMEN COLLECTION:
Curschmann’s Spiral – coiled mucus strands. May be
1. First Morning – Most Preferred
seen macroscopically  Bronchial Asthma
2. 24-hour Sputum – volume measurement
Myelin Globules – May be mistaken as Blastomyces but
3. Throat swab – for pedia patients
has no clinical significance.
4. Sputum Induction
Epithelial Cells/Creola Bodies – cluster of columnar
5. Tracheal aspiration – for debilitated patients.
cells  Bronchial Asthma
Parasites – migrating larva of A. lubricoides, P.
MACROSCOPIC EXAM
westermani, E. histolytica, E. gingivalis, E. granulosus
o VOLUME
Other – neoplastic cells, bacteria and leukocytes
Increased in: Bronchial Asthma, Acute Bronchitis
early Pneumonia
BRONCHIAL ASTHMA:
Decreased In:Bronchiectasis, Lung abcess, edema,
Charcot-Leyden Crystals
gangrene, tuberculosis
Creola Bodies
o ODOR
Curschmann’s Spiral
Foul and Putrid = lung gangrene, necrotizing tumors
Dittrich’s Plug
Sweet = bronchiectasis and tuberculosis
o COLOR
 Colorless/Translucent = pure mucus only BRONCHEOALVEOLAR LAVAGE – collected for
 White/Yellow =  Pus Pneumocystis carinii (jerovecii) in immunocompromised
 Gray =  Pus and Epithelial cells individuals.
 Bright Green/Greenish =  Bile; P. CELLS SEEN IN BRONCHEOALVEOLAR LAVAGE
aeruginosa 56 –
Macrophage Most Predominant
 Red/Bright Red = fresh blood, hemorrhage, 80%
TB, bronchiectasis Interstitial disease,
 Anchovy Sauce/Rusty Brown = Old blood, 1-15% Lymphocytes pulmonary lymphoma,
pneumonia non bacterial infection
 Prune Juice = Pneumonia, chronic lung Cigarette smokers,
cancer <3% Neutrophils bronchopneumonia,
 Olive green/Grass Green = cancer toxin exposure
 Black = inhalation of dust, carbon, charcoal, Hypersensitivity
<1-2% Eosinophils
anthracosis, smoking reactions
 Rusty (with pus) = lobar pneumonia 4-7% Ciliated Columnar Bronchial Epith Cells
 Rusty (without pus) = Congestive heart
failure
 Currant, Jelly-like = K. pneumonia
o CONSISTENCY SWEAT – assayed for the diagnosis of CYSTIC
o Mucoid = asthma, bronchitis FIBROSIS  Autosomal recessive
o Serous/Frothy = lung edema o Metabolic dse that affects mucous secreting
o Mucopurulent = bronchiectasis, glands of the body.
tuberculosis with cavities o Assoc. with pancreatic insufficiency,
repsirstory distress and intestinal
MACROSCOPIC STRUCTURES
obstruction.
Dittrich’s Plug – yellow/gray material, size of a pinhead.
Sweat Sodium and Chloride NV:
Fould odor when crushed. BRONCHITIS and ASTHMA
70 mEq/L = Diagnostic of CF
PNEUMOLITHS – LUNG STONES. Yellow/White calcified
40 mEq/L = Borderline Value
structures or foreign material
BRONCHIAL CAST – tree-like casts. Lobar Pneumonia
GIBSON and COOKE PILOCARPINE IONTOPHORESIS
LAYER-FORMATION: 3 LAYERS
o Pilocarpine + mild current = sweat gland
1st Layer (Top) = Forthy Mucus
2nd layer (Middle) = Opaque, water material stimulation
3rd layer (Bottom) = Pus, Bacteria Tissue o Sweat is tested for Na and Cl.
 1) Bronchoiectasis
 2)Lung Abcess
 3) Gangrene
AMNIOTIC FLUID – Provides cushion for the fetus oSphyngomyelin = serves as control
and permits fetal movement. (contrant production)
 > 2.0 = Mature Lungs
AMNIOTIC FLUID VOLUME – from fetal urine and lung  Reject specimens contaminated with
fluid blood and meconium.
o After the first trimester, fetal urine is the major b. Amniostat-FLM – immunologic test for
contributor to the amniotic fluid volume. phosphatidylglycerol (PG)
o Not affected by blood and meconium
POLYHYDRAMNIOS -  amniotic fluid volume o Production of PG is delayed in diabetic mom
CAUSES:  fetal swallowing of urine c. Foam Stability (Foam Shake Test)
Neural Tube Defects **Amniotic fluid + Ethanol  15 shaking
Stand for 15 mins.
OLIGOHYDRAMNIOS -  amniotic fluid volume (+) Foam/Bubbles = Mature Lungs
CAUSES:  fetal swallowing of urine d. Microviscosity – resence of phospholipids
Membrane Leakage decrease microviscosity.
Urinary Tract Deformities e. Lamellar Body Count – Lamellar Bodies (Type II
SPECIMEN COLLECTION Pneumocytes)
o Method of Collection is called o Responsible for the production of alveolar
_____________________ surfactants.
**up to 30mL is collected in a sterile syringe o >32,000/uL lamellar body count = Adeuqate
FLM
o 2nd Trimester – assess genetic defects (Trisomy f. OD 650 -  Lamellar Bodies =  Optical Density
21/Down Syndrom) o OD of >0.15 = L/S ratio >2.0
o 3rd Trimester – fetal lung maturity (FML), Fetal o (+) Phosphatidylglycerol
Hemolytic Disease (HDN) IV. TEST FOR FETAL AGE
SPECIMEN HANDLING o >2mg/dL Creatinine = 36 weeks (9 months)
o Test for FML – place on ICE (refrigerate/freeze)
o Cytogenetics – room temp (<37C) SUMMARY OF TESTS FOR FETAL WELL-BEING
o HDN – protect from light TEST NV SIGNIFICANCE
Bilirubin Scan Abs at 450 > .025 HDN
AMNIOTIC FLUID vs. MATERNAL URINE -feto protein <2.0 (Multiples of Median) NTD
o CHON and GLUCOSE is present in AMNIOTIC L/S Ratio >2.0 FLM
FLUID not in URINE (Renal Thresholds) Amniostat FLM (+) FLM
o Urine have very high lvls of Creatinine and Urea Foam Stability Index >47 FLM
FERNS TEST Microviscosity (FLM- >55mg/g FLM
TDx)
Specimen (Vag. Fluid)  Slide (Air Dry)  (+) Fern-Like OD 650nm >0.15 FLM
Crystals – AMNIOTIC FLUID Lamellar Body >32,000/mL FLM
Count
COLOR
o Colorless – Normal
HUMAN CHORIONIC GONADOTROPHIN (HCG)
o Blood-Streaked – Traumatic Tap
o Produced by the CYTOTROPOBLAST CELLS of the
o Yellow – HDN (Bilirubin)
o Dark-Green – Meconium placenta
o Dark Red-Brown – Fetal Death o Composed of 2 sub-units
o  = hCG, LH, FSH and TSH
I. TEST FOR HDN o  = unique for hCG
 (O.D. 450) - Normal Absorbance =  at 365nm o Urine hCG spx = 1st morning urine
 at 550nm
** Reported with LILEY GRAPH hCG BIOASSAYS
o Zone 1 = non-affected/mildly affected fetus ANIMAL MODE OF
TEST (+) RESULT
o Zone 2 = moderately affected (requires close USED INJECTION
monitoring) Ascheim- Immature Subcutaneous Formation of
o Zone 3 = Severely affected (requires intervention) Zondeck Female Mice hemorrhagic
follicles and
corpora lutea
II. TEST FOR NEURAL TUBE DEFECT
Friedman Mature Marginal Ear Hyperemic
o Spina Bifida - Anancephaly Virgin Vein uterus and
o SCREENING TEST: Alpha-Feto Protein Female corpora
 in NTD  in Down Syndrome Rabbit hemorrahica
o CONFIRMATORY: Acetylcholinesterase Hogben Female Toad Lymph Sac Oogenesis
South
African
III. TEST FOR FETAL LUNG MATURITY
Clawed Frog
a. Lecithin/Sphyngomyelin Ratio (REFERENCE Galli- Male Frog Subcutaneous Spermatogenesis
METHOD) Mainini Male Toad
o Lecithin = alveolar stability Frank- Immatur e Subcutaneous Ovarian
Berman Female Rats hyperemia
Kupperman Female Rats Intraperitonea Ovarian o For undiluted specimen
l hyperemia o Uses heat to immobilize sperm cells
SEMEN
o For fertility testing SHORT-CUT FOR SPERM CONCENTRATION
o Post-vasectomy semenalysis COMPUTATION
o Forensic analysis (alleged rape) 2 WBC Squares =

COMPOSITION:
5% Spermatozoa 1. Seminiferous Tubules 5 WBC Squares =
2. Epididymis
60-70% Seminal Fluid Seminal Vesciles
20-30% Prostate Fluid Acidic Fluid
ACTUAL SPERM CONCENTRATION COMPUTATION
Contains ACP, Zinc, Citric Acid
and other enzymes Sperm Conc =
For Coagulation and
liquefaction
5% Bulbourethral Fluid Thick alkaline mucus
Neutralize acidity from the
prostatic secretions and the
vagina
SPERM COUNT
 NV: >40 million/ejaculate
SPECIMEN COLLECTION:
 Formula:
1. Abstinence for 2-3 days (>5 days impairs motility)
2. Collect the entire ejaculate:
Sperm count = Sperm conc x Specimen volume
a. Masturbation (the best)
b. Coitus interruptus
SPERM MOTILITY
c. Condom method = use of non-lubricant rubber
 NV: >50% w/i 1hour
or silastic condom.
 Grade: > 2.0
3. Specimen should be delivered to the laboratory
GRADE WHO Criteria
within 1 hour of collection (at RT)
4. Take note of the time of specimen collection, 4.0 a Rapid, Straight-line motility
specimen receipt and liquefaction 3.0 b Slower, some lateral movement
5. Analysis should be done after liquefaction 2.0 b Slow forward, noticeable lateral
(usually 30 to 60C minutes) movement
6. Pending specimen should be kept at 37C 1.0 c No forward progression
0 d No movement
MACROSCOPIC EXAMINATION
Appearance Gray-White, translucent = normal Computer-Assisted Semen Analysis (CASA)
(with musty or bleach odor)  Provides objective determination of both sperm
 white turbidity =  Infection (WBC) velocity and trajectory (direction of motion)
Red/Brown Color = Blood
Yellow =  Abstinence/Contince, Urine SPERM MORPHOLOGY
contamination, medications  NV: Routine: >30% Normal Forms
Volume Normal = 2 to 5 mL Kruger’s Strict Criteria: >14% Normal Forms
 in abstinence o Measure the head, neck and tail using a
 in infertility or incomplete collection micrometer.
Viscosity Normal = Pour in dorplets STAINS: Wright’s, Giema and Papaniculao’s (choice)
 Viscosity =  Motility
Reporting:  Head: Oval (normal) [5um x 3um]
o 0 = Watery o Acrosomal Cap = ½ of the head
o 4 = Gel-Like o Nucleus = 2/3 of the head
pH Normal = 7.2 to 8.0 o Abnormal = poor ovum penetration
 pH = infection  Midpiece: contains mitochondria
 pH =  prostatic fluid  Tail: 45um

VARICOCELE – hardening of the veins that drain the testes

o Most common cuase of male infertility
SPERM CONCENTRATION o Sperm Head = tapered head
 NV: 20 to 160 million/mL
Methods: SPERM VIABILITY
1. Improved Neubauer Counting Chamber Modified Bloom’s Test
o Dilution: 1:20 o Reagents: Eosin and Nigrosin
o Diluents: o Living Sperms: Unstained bluish white (75%)
o Cold Water - Sodium Bicarbonate o Dead Sperms: Red
o Formalin
2. Mackler Counting Chamber SEMEN FRUCTOSE
 o Tested within 2 hours or FROZEN to prevent  VASECTOMY – cutting of vas deferens so that
fructolysis the ejaculate will not contain any sperm cell.
o Screening test: +/- 13 umol/ejaculate  The only concern is the presence or absence of
 Resorcinol Test = (+) Orange-Red Color sperm
 Done 2 months after vasectomy and continued
ANTISPERM ANTIBODY until 2 consecutive monthly specimens show no
o Detected in semen, cervical ucosa or serum specimen
1. Mixed Agglutination Reaction
 Detects the presence of IgG TERMINOLOGY
 Semen + AHG + Latex Particle or IgG Aspermia – No ejaculate
coated RBCs Azospermia – Absence of sperm cells
Necrospermia – Immotile/Dead sperm cells
 Normal: <10% motile sperm attached to
Oligospermia – Decreased sperm concentration
the particle
2. Immunobead Test
 Detects the presence of IgM, IgG and
IgA SYNOVIAL FLUID (Joint Fluid)
 Demonstrate the affected area on the o “Synovial” is latin for EGG.
sperm (head, neck or tail) o Viscous fluid circulating in diarthroses (movable
joint)
CHEMICAL TESTING o FUNCTION:
 Lubricant for joints
Analyte Normal Value  Values
 Reduce friction between bones
Fructose > or = 13  Seminal Fluid  Provides nutrients to the articular cartilage
umol/ejaculate  Lessen shock of joint compression occurring during
Neutral - > or = 20 Epididymis activites such as walking and jogging.
glucosidase mU/ejaculate Disorder SPECIMEN COLLECTION (Arthrocentesis)
Zinc > or = 2.4 o Normally does not clot
umol/ejaculate o VOLUME: <3.5 mL
Citric Acid > or = 52 o INFLAMMATION: >25 mL
 Prostatic Fluid
umol/ejaculate
ACP > or = 200  Tube 1 = Microbio (Heparin)
Units/ejaculate  Tube 2 = Hematology (Sodium Heparin/Liquid EDTA)
*never use powder anticoagulant and lithium heparin—
intrerferes in crystal identification.
MICROBIAL TESTING  Tube 3 = Chemistry and other tests = No Anticoag
o ROUND CELLS  Glucoe Analysis = Sodium Fluoride
o WBCs and spermatids (immature sperm
cells) CLARITY and COLOR
o NV: <1M/mL o Colorless to Pale Yellow = Normal
o >1 million WBCs/mL = infection o Deeper Yellow = Inflammaiton
o >1million spermatids/mL = disruption of o Greenish Tinge = Bacterial Infection
spermatogenesis o Red = Traumatic Tap; hemorrhagic arthritis
o Test for Chlamydia trachomatis, Mycoplasma o Milky = Presence of crystals
hominis and Ureaplasma urealyticum
VISCOSITY
MEDICO-LEGAL TESTS o NORMAL: Forms 4 to 6 cm long string
Test for detection of Semen o TEST: ROPES/MUCIN CLOT TEST (Hyaluronate
1. Microscopic exam Polymerization Test) [Rgnt: 2-5% Acetic Acid]
2. Fluorescence under UV light GRADING:
3. Acid Phospatase determination Good = Solid Clot Low = Friable Clot
4. Glycoprotein p30 = more specific mtd Fair = Soft Clot Poor = No Clot
5. FLORENCE TEST (not specific)
o Test for CHOLINE CELL COUNT
o RGNT: Iodide Crystals + KI WBC Count – most frequently used
o (+) Dark Brown Rhombic Crystals  Diluting Fluid:
6. BARBIERO’s STEST (very specific) 1. NSS with Methylene Blue
o Test for SPERMINE 2. Hypotonic Saline (0.3%)
o RGNT: Saturated Picric Acid + TCA 3. Saline with Saponin
o (+) Yellow Leaf-Like Crystals
7. ABO Blood Grouping **Never use acetic acid as a diluting fluid in WBC count
8. DNA Analysis on SYNOVAL FLUID  Clot formation

POST-VASECTOMY SEMEN ANALYSIS  For very viscous fluid:

 Add pinch of hyaluronidase 0.5mL fluid; or
  Add 1 drop of 0.5% Hyaluronidase in PO 4 electron
buffer per mL of fluid (incubate at 37C for microscopy
5mins)

DIFFERENTIAL COUNT CAUSES OF CRYSTAL FORMATION

RBCs < 2,000/uL 1. Metabolic Disorders
WBCs <200/uL 2. Decreased renal excretion that produce increased
WBC Diff Count blood levels of crystalizing chemicals.
 65% = Monocytes and Macrophages 3. Degeneration of cartilage or bones
 <25% = Neutrophils 4. Medications (Corticosteroids)
 <15% = Lymphocytes
POLARIZING MICROSCOPE – detects presence or
absence of bifringence.
CELLS AND INCLUSIONS IN SYNOVIAL FLUID ** Yellow = Parallel = (-) Bifringence
Cell/Inclusion Description Significance **Blue = Perpendicular = (+) Bifringence
Neutrophil PMN cell Bacterial sepsis
Crystal-induced
inflammation GASTRIC FLUID
Lymphocyte Mononuclear Cells Non-septic
inflamm G cells Gastrin
Macrophage Large mononuclear cells, Normal
may be vacuolated Viral Infections
Synovial Lining Cell Similar to macrophage, Normal Parietal Cells HCl
but may be
multinucleated,
resembling a
mesothelial cell Chief
Pepsinogen Pepsin
LE cells Neutrophils with Lupus Cells
ingested round bodies Erythematosus
Reiter Cells Vacuolated Reiter Syndrome
macrophages with Nonspecific CELLS OF STOMACH:
ingested neutrophils inflamm 1. Parietal Cells – Produces HCl and Intrinsic
RA Cells (Ragocytes) Neutrophil with dark Rheumatoid
cytoplasmic granules Arthritis
Factor
containing immune Immunologic 2. Chief Cells – produces pepsinogen
complexes Inflammation 3. G Cells – produces gastrin
Cartilage Cells Large multinucleated Osteoarthritis
cells SPECIMEN COLLECTION
Rice Bodies Macroscopically Tuberculosis,
resembles POLISHED septic and
o Method of Collection: Gastric Aspiration
RICE. Rheumatoid o Gastric Tubes:
Microscopically show Arthritis 1. Levin Tube = Inserted through nose
collagen and fibrin 2. Rehfuss Tube = Inserted through mouth
Fat Droplets Refractile intracellular Traumatic Injury
and extracellular Chronic
globules inflammation MACROSCOPIC EXAMINATION
Stain (+) with Sudan o COLOR
Hemosiderin Inclusions withihn Pigmented o Pale gray w/ Mucus = Normal
clusters of synovial cells villonodular
o Yellow-Green = Large Amt of Bile
synovitis
o VOLUME
o Fasting = < 50 mL (normal)
CHARACTERISTICS OF SYNOVIAL FLUID CRYSTALS o > 50mL = abnormal fasting specimen
CRYSTAL SHAPE POLARIZATION SIGNIFICANCE o 20-60mL to 120mL = after Ewal’s test meal
Monosodium Needles (-) Bifringence Gout o 45-150mL = after alcohol test meal or histamine
Crystal stimulation
Ca PyroPO4 Rhombic (+) Bifringence Pseudogout
square BASAL ACID OUTPUT (BAO) – total gastric secretion during
rods unstimulated, fasting state
Cholesterol Notched, (-) Bifringence Extracellular MAXIMUM ACID OUTPUT (MAO) – total gastric secretion
rhombic after gastric stimulation
plates
Corticosteroid Flat, (+/-) Injections BAO/MAO RATIO: 0 in Pernicious Anemia |  Zollinger Ellison Syn
variable- Bifringence
shaped
plates GASTRIC STIMULANTS
Calcium Envelopes (-) Bifringence Renal Dialysis Test Meals
Oxalates a. Ewald’s – bread, tea or water
Apatite (Ca Small No Bifringence Osteoarthritis b. Boa’s – oatmeal
PO4) particles; c. Riegel’s – Beef steak or mashed potato
require Chemical Stimulants
 a. Pentagastrin Green Vegetables
b. Histamine Bulky/Frothy Bilie Duct obstruction
c. Histalog (Betazole) Pancreatic disorders
d. Insulin Steatorrhea
Sham Feeding – fictitious feeding (sandwich) Butter-Like Cystic fibrosis
Mucus Colitis
TERMINOLOGIES Blood-Streaked Dystentery
Euchlorhydria – Normal Free HCl mucus Malignancy
Hyperchlorhydria -  Free HCl (Peptic Ulcer) Constipation
Hypochlorhydria -  Free HCl (Stomach Carcinoma) Ribbon-Like Intestinal constriction
Achlorhydria – No Free HCl (Pernicious Anemia) Rice watery Cholera
Pea soup Typhoid
Qualitative Tests for Free HCl Scybalous “Goat Constipation
Dimethylaminoazobenzol – (+) Cherry Red Droppings”
Gunzberg – RGNT: Phloroglucin, Vanillin, Alcohol
= (+) Purple Red Color BRISTOL STOOL CHART
Boas – RGNT: Resorcinol, Cane Sugar, Alcohol TYPE 1 = Separate hard lumps, like nuts (hard to pass)
= (+) Rose Red Color TYPE 2 = Sausage-shaped but lumpy
TYPE 3 = Like a sausage but with cracks on its surface
Quantitative Test for Gastric Acidity TYPE 4 = Like a sausage or snake, smooth and soft
Free HCl Total Acidity Combined HCl
(CHON bound)
TYPE 5 = Soft blobs w/ clear-cut edges (passed easily)
Titrant Sodium Hydroxide Na Sodium TYPE 6 = Fluffy pieces w/ ragged edges, a mushy stool
Hydroxide Hydroxide TYPE 7 = Watery, no solid pieces, entirely liquid
pH Dimethylaminobenzol Phenolphtha Sodium
Indicator (Topfer’s Reagnet) lein Alizarin MICROSCOPIC EXAMINATION
Endpoint Canary Yellow Faint Pink Violet
I. FATS
NV 25 to 50 50 to 75 ⁰ 10 - 15⁰
 _________________ = increased fats in stool
(>6 g/day)
LACTIC ACID – indicative of advanced gastric cancer
 Tests:
TEST FOR LACTIC ACID
1. Screening Test = microscopic examination
Test Reagents Endpoint
Modified FeCl3 + Phenol Yellow
of feces for fat globules
Uffelmann’s 2. Definitive Test = fecal fat determination
Strauss FeCl3 + Ether Yellow FECAL FAT DETERMINATION
Kelling’s FeCl3 Yellow Qualitative Tests
1. Neutral Fat Stain (Triglycerides)
DIAGEX TUBELESS TEST Stool Suspension + 95% Ethanol + Sudan III
 Specimen = Urine 
 Principle: Orange Droplets
o Azure Blue is given by mouth (Neutral Fats/Triglycerides)
o The presence of azure blue in urine  >/= 60 droplets/hpf = Steatorrhea
indicates the presence of free HCl in the
stomach 2. Split Fat Stain (Fatty Acid)
Emulsified stool + 36% AcetOH + Sudan III
FECALYSIS 
Orange droplets
Normal Volume: Around 100-200g stool is passed is
(Fatty Acids)
passed per day.
 Normal = 100 droplets ( <4 um)
o Contains bacteria, cellulose and other foodstuff,
 Slightly Increased = 100 droplets (1-8 um)
gastrointestinal secretions, bile pgments, cells
 Increased = 100 droplet (6-75 um)
from intestinal walls, electrolytes and water.
MACROSCOPIC STOOL CHARACTERISTICS
Quantitative Test
Color/Appearance Clinical Significance
 Van De Kamer Titration
Brown Normal (Urobilin/Stercobilin)
o Gold standard for fecal fat
Black Upper GI Bleeding
o For definitive diagnosis of steatorrhea
Iron therapy
o SAMPLE = 3 days stool (72 hours)
Charcoal
o Titration with NaOH
Bismuth (antacids)
o NaOH value = 1-6g fats/day
Red Lower GI Bleeding
o Steatorrhea = > 6g fats/day
Beets and food coloring
Rifampin
II. MUSCLE
Pale Yellow, White Bile duct obstruction, Barium sulfate
o Creatorrhea = abnormal excretion of muscle
Gray
fibers in feces
Green Biliverdin
o Determination:
Oral antibiotics
 o
The patient should include meat in the diet FECAL CARBOHYDRATES – most valuable in assessing
o
Emulsified stool + 10% Eosin  Coverslip cases of infant diarrhea (Ex. Lactose intolerance)
and stand for 3mins  Count the number FECAL CARBOHYDRATE DETERMINATION
of undigested fibers (HPF) 1. Clinitest 2. pH
 Completely Digested = No striations  Test for reducing  Normal stool pH =
 Partially digested = striation in one direction sugars 7.0 to 8.0
 Undigested = striation in both directions  >0.5g/dL  CHO disorder = pH
**ABNORMAL > 10 undigested muscle fiber carbohydrate <5.5
 Biliary obstruction intolerance
 Cystic fibrosis CEREBROSPINAL FLUID
o 3rd major body fluid
III. FECAL LEUKOCYTE o Functions:
 >/= 3 neutrophils/HPF = invasive condition o Supply nutrients to the nervous system
Diarrhea w/ WBCs Diarrhea w/o WBCs o Remove metabolic wastes
Salmonella Toxin producing: o Produce mechanical barrier to cushion the brain
Shigella Staphylococcus aureus and spinal cord against trauma
Yersinia Vibrio cholera MENINGES – lines the brain and spinal cord
Enteroinvasive E. coil Virus 3 LAYERS:
Campylobacter Parasites 1: Dura Mater (Outer Layer) – lines the skull and
Determination: vertebral canal
1. Wet Preparation = Stool + Loefflers 2: Arachnoid Mater (Spider-Like) – filamentous inner
Methylene Blue membrane
2. Dried Preparation = Stool + Wright’s/Gram’s  Subarachnoid space – where CSF flows
Stain 3. Pia Mater (Innermost) – lines the surface of the
3. Lactoferrin Latex Agglutination Test brain and spinal cord.
 Lactoferrin = 2⁰ granules of neutrophils
 (+) Invasive bacterial pathogen CHOROID PLEXUS – for CSF production
o 20mL/hr =rate of CSF production
FECAL OCCULT BLOOD TEST ARACHNOID VILI/GRANULAITON – reabsorbs CSF (back
Notes Occult = Hidden
Screening Test for Colorectal cancer to circulation)
Significant = >2.5mL blood/150g stool BLOOD BRIAIN BARRIER – protects brain from other
Sample = center portion of the stool chemical and substances circulating in the blood that
Principle Pseudoperoxidase activity of Hgb can harm the brain tissue.
H2O2 + Guiac  Oxidized Guiac (Blue) + H2O
o Disruption of BBB allows WBCs, proteins and
Chromogens 1. Benzidine
2. Guiac other chemicals to enter the CSF (Ex.
3. O-Toluidine Meningitis, Multiple Sclerosis)
False (+) Dietary pseudoperoxidase
 Red Meat (Avoid 3 days) CSF COLLECTION AND HANDLING (lumbar puncture) -
 Melon, broccoli, cauliflower, horseradish (Avoid 3
Between 3rd, 4th or 5th lumar vertebrae (up to 20mL may
days)
 Aspirin and NSAIDs (Avoid 7 days) be collected)
False (-) Reducing Agents
CSF TUBE 1 = Chem/Sero (frozen)
APT TEST PROCEDURE CSF TUBE 2 = Bacte (RT)
Emulsified Stool  Centrifuge  Add 1% NaOH to CSF TUBE 3 = Hema/Cell Count (Refrigerate)
Supernatant CSF TUBE 4 = Microbio/Sero
Pink Solution = (+) Fecal blood (Hgb F) *if 1 tube only = Bactte  Hema  Chem/Sero
Yellow-Brown Supernatant = (+) Maternal blood (Hgb A)
APT TEST PROCEDURE – differentiates fetal blood and Normal Volume:
maternal blood ADULTS: 90 to 150mL (Stras) [140 to170mL old Stras]
 Specimen = Infant stool and vomitus NEONATES: 10 to 60mL
Note:
 Hemoglobin F is alkali-resistant APPEARANCE
 Hemoglobin A is denatured by NaOH Appearance Significance
Crystal Clear Normal (<200/uL WBC, <400/uL RBC)
Hazy/Turbid  WBC (>200/uL)
X-RAY FILM TEST PROCEDURE
Milky/  RBC (> 400/uL)
Emulsified Stool  X-ray film CLoudy  Lipids and CHON
**Clearing of Film = (+) Trypsin (+) Microorganisms
**No Clearing of Film = (-) Trypsin Xanthochro Hgb degradation
X-RAY FILM TEST mic Pink =  Oxyhemoglobin
o Detects trypsin enzyme (Pink/ Yellow = OxyHgb to Bilirubin
o (-) Trypsin = Seen in Cystic Fibrosis Yellow/ Orange =  Hemolysis
Orange) Other cause:  Carotene,  Melanin, Rifampin
Bloody  RBC (>6,000/uL)
 Traumatic Tap PREDOMINANT CELLS IN CSF
o Uneven blood distribution among the o Predominant = Lymphocyte and Monocytes
tubes o Occasional = Neutrophils
o May clot o ADULT RATIO: 70 Lympho:30 Mono
o Clarifies upon spinning
o NEONATES: 70 to 80 Mono: 20 to 30 Ly
o (-) Erythrophage
Intracranial Hemorrhage
Pleocytosis – increased number of normal cells in CSF
o Even blood distribution among the
tubes TYPE OF CELL CLINICAL SIGNIFANCE
o No fibrinogen Lymphocytes Normal
o Xathochromia Monocytes Viral tubercular and fungal
o (+) Erythrophage meningitis
Oily Radiographic contrasts media Multiple sclerosis
Clotted Protein and Clotting Factor Neutrophil Bacterial Meningitis
Pellicle Tubercular Meningitis Early viral, tubercular and
fungal meningitis
CSF CELL COUNT – any cell count should be performed Macrophage RBC in spinal fluid
IMMEDIATELY Contrast media
o WBCs and RBCs begin to lyse w/i 1hr. Blast Forms Acute Leukemia
o 40% WBCs disintegrate w/i 2hrs Lymphoma Cells Disseminated Lymphoma
Plasma Cells Multile Sclerosis
1. WBC Count – routinely performed in CSF Lymphocyte reactions
NV: ADULT – 0.5 WBC’s/uL Ependymal, Choroidal and Diagnostic procedures
NEONATE – 0 – 30 WBC/s/uL Spindle-shaped Cells
CSF DILUTION – 3% Acetic Acid w/ Methylene Blue Malignant Cells Metastatic carcinomas
APPEARANCE DILUTION Primary CNS Carcinoma
Clear Undiluted
Slightly Hazy 1:10 CSF PROTEINS
Hazy 1:20  Normal Value
Slightly Cloudy 1:100  Adults: 15 to 45 mg/dL
Cloudy/Slightly Bloody 1:200  Infants: 150 mg/dL
Bloody/Turbid 1:10,000  Immaure: 500 mg/dL
 Increased In:
2. RBC Count – done only in cases of traumatic tap  Damage to BBB (Meningitis, Hemorrhage)
 Correction is made for WBC and total CHON  Production of Igs in CNS (Multiple Sclerosis)
concentration:  Decreased In:
o Subtract 1 WBC for every 700 RBCs seen  CSF Leakage
o Subtract 1mg/dL Total CHON for every
1,200 RBCs/uL  Major CSF Protein: Albmin,
 2nd most: Pre-Albumin
CSF DIFFERENTIAL COUNT
 Performed on stained smear  -globulin – Haptoglobin and ceruloplasmin
 Specimen should be concentrated before  -globulin - -transferrin
smearing by using the following methods o Carbohydrate-deficient transferrin
1. Cytocentrifugaiton o Found in CSF but not in serum
2. Centrifugation  -globulins – IgG and IgA
3. Sedimentation  NOT NORMALLY FOUND IN CSF: lgM, Fibrinogen
4. Filtration and Lipids

CYTOCENTRIFUGE – for concentration of cells not CSF PROTEIN DETERMINATION

separation. A. TOTAL PROTEINS:
 Fluid is poured in a conical chamber 1. Turbidimetric
 Cells are forced into a monolayer w/i 6mm a. TrichloroAcetic Acid (TCA) – preferred
diameter circle on the slide method
 Addition of albumin:  Precipitates BOTH Alb and
o Increases cell recovery Globulins
o Decreases cellular distortion b. Sulfosalycylic Acid
HORIZONTAL CENTRI ANGLE HEAD  Precipitated Alb only; to
Vertical (Not spinning) to Fixed Angle precipitate Globulins, add Na
Horizontal (Spinning) Sulfate.
 Air friction =  Air  Air Friction =  Air 2. Dye-Binding (Coomassie Brilliant Blue)
resistance Resistance o CHONs bind to dye  Dye turn RED TO
Speed: 1,650 g 9,000 g BLUE
o Increased Blue color = Increased Protein
 1. Serum LDH – 2>1>3>4>5
B. PROTEIN FRACTIONS Flipped= 1>2 (AMI)
1. CSF/SERUM ALBUMIN INDEX: assessment of 2. CSF LDH
integrity of BBB NORMAL: 1>2>3>4>5
 Normal Value: <9 Neurobiological abnormalities: 2>1
 Abnormal: >9 Bacterial Meningitis: 5>4>3>2>1
- Correlated to degree of damage
- Index of 100 = COMPLETE BBB damage LDH ISOENZYMES IN CSF:
2. IgG INDEX LD 1 and 2: Brain tissue
o Assessment of conditions w/ IgG production w/i LD 2 and 3: Lymphocytes
CNS (Multiple Sclerosis) LD 4 and 5: Neutrophils
o (CSF IgG/Serum IgG)/(CSF/Serum Alb Index)
 Normal Value: <0.77 MAJOR LABORATORY RESULT FOR DIFFERENTIAL
 Abnormal Value: >0.77 DIAGNOSIS OF MENINGITIS
 Indicative of IgG production w/i CNS Bacetrial VIral Tubercular Fungal
WBC 
ELECTROPHORESIS    Lympho Lympho
o Done in conjunction with serum electrophoresis Neutro Lympho and Mono and
o For the detection of OLIGOCLONAL BANDS (in Mono
the -region (Indicative of IG production) CHON    
o The presence of 2 or more oligoclonal bands in Glu  Normal  
CSF bu not in serum is valueable for the Lactate  Normal  
diagnosis of MULTIPLE SCLEROSIS Others (+) Gram Agents: Agent: MTB Agent: C.
stain Enterovirus (+) AFB Stain neoformans
MULTIPLE SCLEROSIS : Demyelinating (+) Culture Poliovirus (+)
Disorder (+) Lymulus Echovirus Pellicle/weblike (+) GS =
Lysate Test Coxackie Clot formation
Findings: virus after 12-24
Class
 (+) Anti-Myelin Sheath autoantibody Starburst
hour patter
 (+) Oligoclonal band in CSF but not in refrigetation (+) India Ink
Serum (+)
 (+) Myelin basic protein Immunologic
  IgG index tests
MYELIN BASIC PROTEIN –protein component of lipid-
protein complex that insulates the nerve fibers
o Presence of MBP in CSF indicates destruction of LYMULUS LYSATE TEST – detects gram negative
myelin sheath bacterial endotoxin in body fluid and surgical
o Used to monitor the course of multiple sclerosis instruments
REAGENT: Blood of horshoe crab
CSF GLUCOSE Principle:
Determination Done in conjunction with blood  In presence of endotoxin, the amoeboctes
glucose (blood specimen mut be (WBCs) will release lysate
collected 2 hours before spinal  (+) Clumping/ Clot Formation
tap)
Normal Values 60 to 70% of Blood glucose (50 to AGENTS FOR BACTERAIL MENINGITIS:
80mg/dL) Age Group Causative Agents
Increased in Due to increased plasma glucose Birth to 1 month S. agalactiae
Decreased in Bacterial meningitis 1 month to 5 y/o H. influenza
Tubercular eningitis 5 to 29 y/o N. meningitides
Fungal meningitis >29 y/o S. pneumonia
Normal in Viral meningitis Infants, elderly, L. monocytogenes (walang
immunocompromised pinipili)
CSF LACTATE
Normal Value 10 to 22 mg/dL SEROUS FLUID
Increased In Bacterial Meningitis(>35mg/dL) o Fluid between parietal and visceral membranes
Tubercular Meningitis (>25mg/dL) o Function: provides lubrication between 2
Fungal Meingitis (>25mg/dL) membranes as the surface move against each other.
Normal In Vral Meningitis
EFFUSION – accumulation of fluids between the
CSF GLUTAMINE membranes. May be Exudate or transudate
Product of ammonia and -ketoglutarate
NV: 8 to 18mg/dL Transu Exu
Increased in: Disturbance of consciousness (coma) Appearance Clear Cloudy
REYEs Syndrome Fluid:Serum CHON ratio < 0.5 > 0.5
Fluid:Serum LD Ratio < 0.6 > 0.6
CSF ENZYMES – Lactate Dehydrogenase (LDH)
WBC Count < 1,000/uL .> 1,000/uL 3. Mesothelial Cells – Normal and reactive forms have
Spontaneous clotting No Possible no clinical significance. Decreased in TB
Pleural Fluid: Cholesterol < 45-60 .> 60 4. Plasma Cells – TB
Pleural Fluid:Serum < 0.3 .> 0.3 5. Malignant Cells – Pimary adenocarcinoma, small cell
Cholesterol ratio carcinoma
Pleural Fluid Bilirubin Ratio < 0.6 .> 0.6 Tumor Markers
Serum-Ascites Albumin > 1.1 < 1.1 1. CEA (Carcinoembryonic Antigen) – Colon Cancer
Gradient 2. CA 125 – Ovarian/Metastatic Uterine Cancer
Glucose   3. CA 15-3, CA 549 – Breast Cancer
Rivalta’s Test (-) (+) 4. CYFRA-21-1 (Cytokeratin FRAgment) – Lung
Cancer
RIVALTA’S TEST (Seroamucin Clot Test) - Differentiates
exudates from Transudates Significance of Tests for Pleural Fluid
1. Glucose -  in RA, Purulent Infection
AcetOH + Fluid  Heavy Precipitation = EXUDATE 2. Lactate -  in Bacterial infection
3. Triglycerides -  Chylous effusions
METHOD OF SAMPLE COLLECTION (3P’s) 4. pH -  pneumonia not responding to antibiotics
Normal Apearance = Clear, Pale Yellow  Esophageal Rupture
 Pleaural Fluid – Thoracentesis 5. Adenosine Deaminase (ADA) –  Tuberculosis,
 Pericarcial Fluid – Pericardiocentesis malignancy
6. Amylase -  Pancreatitis, Esophageal Rupture and
 Peritoneal Fluid – Paracentesis
malignancy

PLEURAL FLUID PERICARDIAL FLUID

Appearance Significance
SIGNIFICANCE OF TESTING
Clear, Pale-Yellow Normal
Appearance Significance
Turbid, White Microbial Infection (TB)
Clear, Yellow Normal, Transudate
Brown Rupture of amoebic liver abcess
Blood Streaked Infection, Malignancy
Black Aspergillosis
Grossly bloody Cardiac Puncture,
Viscous Malignant mesothelioma
anticoagulant medication
(Increased hyaluronic acid)
Diff Count
Milky Chylous material
Neutrophils Bacterial endocarditis
Pseudochylous material
Malignant Cells Metastatic carcinoma
Cloudy Hemothorax
Hemorrhagic effusion Test
Gram Stain Bacterial endocarditis
MILKY PLEURAL FLUID AFB Tubular effusion
CHYLOUS PSEUDOCHYLOUS ADA Tubercular effusion
EFFUSION EFFUSION
Cause Thoracic Duct Chronic PERITONEAL/ASCITIC FLUID
Leakage Inflammation APPERANCE
Appearance Milky/White Milky/Green Clear, Pale Yellow Normal
Tinge/ Gold Paint
Turbid Microbial infection
WBC  Lymphocytes Mixed Cells
Green Gall bladder, pancreatic
Cholesterol Absent Present disorders
Crystals
Blood-Streaked Trauma, Infection,
Triglycerides > 110 mg/dL < 50 mg/dL Malignancy
Sudan III (+++) (-) or weakly (+) Milky Lymphatic trauma and
blockage
BLOODY PLEURAL FLUID WBC COUNT: < 500/uL NORMAL ( in Bacterial
HEMOTHORAX HEMORRHAGIC infection and Cirrhosis)
EFFUSION TESTS SIGNIFICANCE
Distribution of Uneven Even Peritoneal Lavage >100,000 RBC/uL indicates
Blood TRAMA (intra-abdominal
Hematocrit Pleural Fluid > Pleural Fluid Hct bleeding)
Whole Blood Hct < Whole Blood CEA Malignancy of GI Origin
(> 50%) Hct (< 50%) CA 125 Malignancy of Ovarian
Significant Cells Origin
1. Neutrophil – Pneumonia, pancreatitis, pulmonary Glucose  Tubercular Peritonitis,
infarction malignancy
2. Lymphocyte – TB, viral infection, autoimmune
Amylase  Pancreatitis, GI
disorders, malignancy
perforation
BUN/Creatinine Ruptured/Puncture
 bladder
GS/CS Bacterial peritonitis
AFB Tubercular peritonitis
ADA Tubercular peritonitis

PSAMMOMA BODIES – contains striation of collagen-

like material
- Seen in benign conditions and associated with
ovarian and thyroid carcinoma