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URINALYSIS
Nephron Basic structural unit of kidney
1M/kidney
Urethra F: 3-4 cm
M: 20 cm
Urine formation (order) Glomerulus à Bowman’s capsule à PCT à Loop of Henle à DCT à CD
PCT 65% of reabsorption
ADH Regulate H2O reabsorption in DCT and CD
Urine composition 95-97% H2O
3-5% solids
60g TS in 24 hrs
35g: Organic = Urea (major)
25g: Inorganic = Cl (#1) > Na+ > K+
Glomerular Filtration
Clearance tests Evaluate glomerular filtration
1. Urea clearance
2. Creatinine clearance = most common
3. Inulin clearance = gold standard
4. Beta2-microglobulin
5. Radioisotopes
Creatinine clearance Formula:
Cc = U x V x 1.73
P A
Normal values:
M = 107-139 mL/min
F = 87-107 mL/min
Tubular Reabsorption
Tubular Reabsorption 1st function to be affected in renal disease
Concentration tests Evaluate tubular reabsorption
Fishberg test (Old) Patient is deprived of fluid for 24hrs then measure urine SG
(SG ≥ 1.026)
Mosenthal test (Old) Compare day and night urine in terms of volume and SG
Specific Gravity (New) Influenced by # and density of particles in a solution
Osmolarity Influenced by # of particles in a solution
Principle: Freezing point depression
- 1 Osm or 1000 mOsm/kg of H2O will lower the FP of H2O (0’C) by 1.86’C
- âFP = áOsm
Example:
Determine Osm in mOsm/kg
Temp. = -0.90’C
Solution:
1000 mOsm/kg = _ _x____
-1.86’C -0.90’C
x = 484 mOsm/kg
Tubular Secretion and Renal Blood Flow
PAH test p-aminohippuric acid
PSP test Phensulfonphthalein test
Obsolete, results are hard to interpret
Methods of Collection
Midstream/Catheterized Urine culture
Suprapubic aspiration Anaerobic urine culture
3 glass technique For detection of prostatic infection
1. 1st portion of voided urine
2. Middle portion of voided urine: Serves as control for kidney and bladder infection
-If (+), result for #3 is considered invalid
3. Urine after prostatic massage
Compare WBC and Bacteria of specimen 1 and 3
Prostatic infection: 1 < 3 (10x)
Pediatric specimen Wee bag
Drug Specimen Collection Chain of custody: step by step documentation of handling and testing of legal specimen
Required amount: 30-45 mL
Temperature (urine): 32.5-35.7’C (w/in 4 mins)
Blueing agent à Toilet bowl (to prevent adulteration)
Types of Urine Specimen
Occasional/Single/Random Routine
Qualitative UA
24 hr 1st voided urine à discarded
w/ preservative
Ex. 8AM à 8AM
12 hr Ex. 8AM à 8PM
Addis count: measure of formed elements in the urine using hemacytometer
Afternoon (2PM-4PM) Urobilinogen (alkaline tide)
4 hr Nitrite determination (1st morning/4 hr)
NO3 à NO2 = (+) UTI
1st morning Pregnancy test (hCG)
Ideal specimen for routine UA
Most concentrated and most acidic = preservation of cells and casts
Fasting/2nd morning Glucose determination
2nd voided urine after a period of fasting
Changes in Unpreserved Urine
Decreased
Clarity Bacterial multiplication
Precipitation of AU/AP
Glucose Glycolysis
Ketones Volatilization
Bilirubin Photooxidation
Urobilinogen Oxidized to urobilin
RBC/WBC Disintegrate in alkaline urine
Increased
pH Urea ---(Urease)---> NH3
Bacteria Multiplication
Odor Urea ---(Urease)---> NH3
Nitrite Bacterial multiplication
Differentiate contamination from Contamination: á Bacteria
true infection True infection: á Bacteria and WBCs
Preservation
Refrigeration 2-8’C
á SG (hydrometer/urinometer)
Precipitate AU/AP
Formalin Addis count
Boric acid Urine culture
Bacteriostatic to contaminants
Sodium fluoride Glucose
Sodium benzoate/ Substitute for sodium fluoride
Benzoic acid
Saccomanno’s fixative 50% ethanol + carbowax
Cytology (50mL urine)
Physical Examination of Urine
Volume NV:
24 hr = 600-1200 mL
Ave (24 hr) = 1200-1500 mL
Night: Day ratio = 1:2 to 1:3
Routine UA Vol = 10-15 mL (Ave: 12 mL)
-15 mL: for urinometry
-physical, chemical, microscopic exam
Polyuria á Urine volume
-Diabetes Mellitus: á vol, á SG
-Diabetes Insipidus: á vol, â SG
Oliguria Calculus/kidney tumors
Dehydration
Anuria Complete cessation of urine flow
Nocturia >500mL w/ SG <1.018
Pregnancy
Urine color Roughly indicates the degree of hydration
Should correlate w/ urine SG
Soluble w/ heat AU
Uric acid
Soluble w/ dilute acetic acid RBCs
AP
Carbonates
Insoluble in dilute acetic acid WBCs
Yeasts
Spermatozoa
Bacteria
Soluble in ether Lipids
Lymph fluid
Chyle
Specific Gravity
SG Density of solution compared w/ density of similar volume of distilled H2O at a similar
temperature
NV = 1.003-1.035 (random)
SG <1.003 = not a urine except DI
Refractometer (TS meter) Based on refractive index:
RI = _light velocity in air_
light velocity in soln
Compensated to temperature (15-38’C)
Corrections:
a. 1g/dL glucose: (-0.004)
b. 1g/dL protein: (-0.003)
Calibrations:
a. Distilled H2O = 1.000
b. 5% NaCl = 1.022 ± 0.001
c. 9% Sucrose = 1.034 ± 0.001
Urinometer Requires temperature correction
a. á 3’C calibration temperature (20’C) = (+0.001)
b. â 3’C calibration temperature (20’C) = (-0.001)
Requires correction for glucose and protein (Rf/U)
Rf < U by 0.002 Refractometer reading is lower than the urinometer reading by 0.002
Urinometer calibration K2SO4 solution: 1L H2O + 20.29g K2SO4
SG = 1.015
Isosthenuria SG = 1.010 (Glomerular filtrate)
Hyposthenuria SG < 1.010
Hypersthenuria SG > 1.010
Urine Odor
Aromatic/Odorless Normal
Ammoniacal Urea ---(Urease)---> NH3
Ex. UTI (Proteus: á urease)
Fruity, sweet DM (Ketones)
Rotten fish/Galunggong Trimethylaminuria
Sweaty feet Isovaleric acidemia
Mousy Phenylketonuria
Cabbage Methionine malabsorption
Caramelized sugar, curry MSUD
Bleach Contamination
Sulfur Cystine disorder
Principle (Rgt strip) Indoxyl carbonic acid ester + Diazonium salt ---(LE)---> Indoxyl + Acid indoxyl
----------> (+) Purple
Strip can detect even lysed WBCs
Reading Time (Reagent Strips)
30 seconds Glucose
Bilirubin
40 seconds Ketones
45 seconds SG
60 seconds “PPBUN”
pH
Protein
Blood
Urobilinogen
Nitrite
120 seconds Leukocytes
Vitamin C (Ascorbic acid) 11th reagent pad
Reducing property
False (-) rgt strip: “BB LNG”
-Blood
-Bilirubin
-Leukocytes
-Nitrite
-Glucose
Rgt: Phosphomolybdate
Phosphomolybdate + Vitamin C (≥5 mg/dL) --------> (+) Molybdenum blue
False-positive False-negative
Blood Strong oxidizing agents High specific gravity/crenated cells
Bacterial peroxidases Formalin
Menstrual contamination Captopril
High concentration of nitrite
Ascorbic acid >25 mg/dL
Unmixed specimens
Bilirubin Highly pigmented urines, phenazopyridine Specimen exposure to light
Indican (intestinal disorders) Ascorbic acid >25 mg/dL
Metabolites of Lodine High concentrations of nitrite
Urobilinogen Porphobilinogen Old specimens
Indican Preservation in formalin
p-aminosalicylic acid
Sulfonamides
Methyldopa
Procaine
Chlorpromazine
Highly pigmented urine
Nitrite Improperly preserved specimens Nonreductase-containing bacteria
Highly pigmented urine Insufficient contact time between bacteria and nitrate
Lack of urinary nitrate
Large quantities of bacteria converting nitrite to
nitrogen
High concentrations of ascorbic acid
High specific gravity
Leukocytes Strong oxidizing agents High concentrations of protein, glucose, oxalic acid,
Highly pigmented urine, nitrofurantoin ascorbic acid, gentamicin, cephalosporins,
tetracyclines
Sources of error:
-Yeasts
-Oil droplets
-Air bubbles
-CaOx crystals
♫ Remedy: add 2% acetic acid
-RBCs: lysed
-Other cells: intact
WBCs NV = 0-5 or 0-8/hpf
Glitter cells (Hypotonic urine)
-Granules swell
-Brownian movement
>1% eosinophils: significant
-á Drug-induced allergic reaction
-á Inflammation of renal interstitium
Addis count Quantitative measure of formed elements of urine using hemacytometer
Specimen: 12 hr urine
Preservative: Formalin
NV:
a. RBCs: 0-500,000/12 hr urine
b. WBCs: 0-1,800,000/12 hr urine
c. Hyaline Casts: 0-5000/ hr urine
Squamous epithelial cells Largest cell in the urine sediment
From linings of vagina, female urethra and lower male urethra
♫ Variation: Clue cells:
-EC w/c are studded w/ bacteria (bacterial vaginosis)
-Whiff/Sniff test: vaginal discharge + 10% KOH à Fishy amine-like odor
-Culture: G. vaginalis = HBT medium
Transitional epithelial cells Spherical, polyhedral, or caudate w/ centrally located nucleus
(Urothelial cells) Derived from the linings of the renal pelvis, ureter, urinary bladder, male urethra
(upper portion)
Not clinically significant in small numbers
Renal tubular epithelial cells Rectangular, polyhedral, cuboidal or columnar w/ an eccentriac nucleus, possibly
bilirubin stained or hemosiderin laden
From nephron:
-PCT: rectangular, columnar/convoluted
-DCT: round/oval
>2 RTE/hpf: tubular injury
Oval fat body Lipid containing RTE cells
Lipiduria (Ex. nephrotic syndrome)
Cholesterol: Maltese cross
Bubble cells RTE cells w/ nonlipid containing vacuoles
Acute tubular necrosis
Yeast C. albicans (DM, vaginal moniliasis)
T. vaginalis Flagellate w/ jerky motility
Pingpong disease
S. haematobium “Hematuria”
Specimen: 24 hr unpreserved urine
E. vermicularis Most common fecal contaminant
Casts (Cylindruria) Formed in the DCT and CD
♫ Tamm-Horsfall protein (Uromodulin)
-Major constituent
-Glycoprotein secreted by RTE cells of DCT and CD
Hyaline casts NV = 0-2/lpf
Beginning of all types of casts (prototype cast)
a. Physiologic:
- Strenuous exercise (HC, GC, RC)
- Heat
b. Pathologic:
- GN
- PN
- CHF
RBC casts Bleeding w/in the nephron
a. GN
b. Strenuous exercise (HC, GC, RC)
WBC casts Inflammation w/in the nephron
Differentiates upper UTI (pyelonephritis, w/ cast) from lower UTI (cystitis, no cast)
To differentiate from EC cast:
1. Phase contrast microscopy
2. Supravital stain
Seen in:
-PN
-AIN
Bacterial casts Pyelonephritis
Epithelial cell casts Renal tubular damage
Advanced tubular destruction
Coarse/Fine granular casts Formed from the disintegration of cellular cast
GN
PN
Strenuous exercise (HC, GC, RC)
Fatty casts Nephrotic syndrome: lipiduria
Not stained by Sternheimer-Malbin
Waxy casts Final degenerative form of all types of casts
Stasis of renal flow
Chronic renal failure
Brittle, highly refractile, w/ jagged ends
Broad casts “Renal failure casts”
Extreme urine stasis
Widening and destruction of tubular walls
Any type of cast can be broad
Sediment preparation Urine à Centrifuge: 400 RCF for 5 mins à Decant à Remaining: 0.5mL/1.0mL
Urine sediment: 20µL (0.02 mL)
-10 lpf
-10 hpf
-Reduced light
RCF 1.118 x 10-5 x radius (cm) x (rpm)2
Urine Crystals
Amorphous Urates Yellow-brown granules
(Normal) Pink sediment (Uroerythrin)
(pH: acid) Mistaken as cystine crystals
Rhombic, wedge, rosette, hexagonal, four-sided plate (whetstone)
Lemon-shaped (Henry)
á Lesch-Nyhan syndrome: orange sands in diaper
á Gout
á Chemotherapy
Calcium Oxalate 1. Weddelite = dihydrate
(Normal) -Envelope/pyramidal
(pH: acid/alkaline/neutral) 2. Whewellite = monohydrate
-Oval, dumbbell
-Ethylene glycol poisoning (antifreeze agent)
Most renal stones consist of CaOx
Amorphous Phosphates White precipitate
(Normal) Granular appearance
(pH: alkaline/neutral) á After meal (alkaline tide)
Ammonium Biurate Yellow-brown
(Normal) Thorny apples
(pH: alkaline) Old specimen: due to the presence of urea-splitting bacteria
Triple Phosphate A.k.a. Magnesium ammonium phosphate
(Normal) Coffin lid, “Struvite”, staghorn appearance
(pH: alkaline) Presence of urea-splitting bacteria
Calcium Phosphate Colorless, flat rectangular plates or thin prisms often in rosette formation
(Normal) Rosettes may resemble sulfonamides
(pH: alkaline/neutral) -To differentiate: CaPO4 dissolves in acetic acid
1. Calcium Phosphate = Apatite
2. Basic Calcium Phosphate = Hydroxyapatite
3. Calcium Hydrogen Phosphate = Brushite
Calcium Carbonate Small and colorless
(Normal) Dumbbell or spherical shapes
(pH: alkaline) Acetic acid: (+) Effervescence
Cystine Colorless hexagonal plates
(Abnormal) Cystinuria
(pH: acid)
Cholesterol Rectangular plate w/ notch in one or more corners
(Abnormal) Staircase pattern
(pH: acid) Lipiduria (Nephrotic syndrome)
Resemble crystals of RCM, to differentiate
a. Patient history
b. Correlate w/ other UA results
c. RCM: áSG by refractometer ≥1.040
Tyrosine Colorless to yellow needles
(Abnormal) Liver disease (more common)
(pH: acid/neutral) (+) Nitroso-naphthol
Leucine Yellow-brown spheres w/ concentric circles and radial striations
(Abnormal) Liver disease
(pH: acid/neutral)
Bilirubin Clumped needles or granules w/ yellow color
(Abnormal) (+) Diazo reaction
(pH: acid) Liver disease
Sulfonamide Colorless to yellow brown
(Abnormal) Deposits in nephrons
(pH: acid/neutral) Tubular damage
Needles, sheaves of wheat, rosette (res. CaPO4 rosette)
♫ Lignin test:
Newspaper = urine + 25% HCl à (+) Yellow orange color
Renal Diseases
Cystitis Inflammation of urinary bladder
Infection
WBCs, RBCs, bacteria
NO CAST
Urethritis Inflammation of urethra
WBCs, RBCs
Usually NO BACTERIA on routine UA
a. Male: GS of urethral exudates [Gram (-) diplococcic]
b. Female: pelvic exam for vaginitis and cervicitis
Glomerulonephritis Inflammation of the glomerulus
Immune-mediated
RBCs, WBCs, RBC CASTS, WBC casts, hyaline and granular casts
Pyelonephritis Infection of renal tubules
WBCs, RBCs, bacteria, RBC casts, WBC CASTS, hyaline and granular casts
Acute Interstitial Nephritis Infection of the renal interstitium
RBCs, WBCs, WBC casts, NO BACTERIA
Renal carcinoma 1’ = RCC
2’ = Transitional CC
RBCs and WBCs
Nephrotic syndrome Massive proteinuria and lipiduria
a. Serum (Chemistry)
-â Albumin, alpha1, beta and gamma globulins
-á alpha2 (AMG)
b. Urine (CM)
-á Albumin, alpha1, beta and gamma globulins
-(-) alpha2 (AMG)
-Oval fat bodies, fatty and waxy casts
Telescoped sediments Simultaneous appearance of the elements of acute/chronic GN and nephrotic syndrome
á Cells and Casts
a. Lupus nephritis
b. SBE
UTI E. coli = 90% cases of UTI
S. saprophyticus = UTI among sexually active young females
G. vaginalis = bacterial vaginosis
S. pyogenes = AGN and ARF
Viridans Streptococci = SBE
Rapidly progressive Deposition of immune complex from systemic immune disorders on the glomerular
(Crescentic) GN membrane
Goodpasture syndrome Attachment of cytotoxic antibody to glomerular and alveolar basement membrane
Wegener’s granulomatosis Antineutrophilic cytoplasmic autoantibody
Henoch-Schönlein purpura Occurse in children following viral respiratory infection
Decrease in platelets disrupts vascular integrity
Membranous GN Thickening of the glomerular membrane following IgG immune complex deposition
Membranoproliferative GN Cellular proliferation affecting the capillary walls or the glomerular basement membrane
Chronic GN Marked decrease in renal function resulting from glomerular damage precipitated by
2. Renal type
N-AA in blood
Impaired tubular reabsorption of AA
Ex. Cystinuria (COLA), Fanconi’s syndrome
Phenylalanine-Tyrosine Disorders
Phenylalanine (-)
PAH PKU Phenylpyruvic acid
Tyrosine
Tyrosine transaminase (-)
p-Hydroxyphenylpyruvic acid Tyrosinemia Tyrosyluria:
p-Hydroxyphenylpyruvic acid oxidase p-OHPPA
Homogentisic acid (-) p-OHPLA
Homogentisic acid oxidase Alkaptonuria
Maleylacetoacetic acid Homogentisic acid
Fumarylacetoacetic acid
Patient preparation
Specimen collection, handling and storage
Analytical Factors Reagents
Instrumentation and equipment
Testing procedure
QC
Preventive maintentance
Access to procedure manuals
Competency of personnel performing the tests
Microscopic Quantitations
EC (lpf) Crystals (hpf) Bacteria (hpf) Mucous threads
None 0 0 0 -
Rare 0-5 0-2 0-10 0-1
Few 5-20 2-5 10-50 1-3
Moderate 20-100 5-20 50-200 3-10
Many >100 >20 >200 >10
Casts (lpf) None = 0
Numerical ranges = 0-2/2-5/5-10/>10
RBCs (hpf) None = 0
Numerical ranges = 0-2/2-5/5-10/10-25/25-50/50-100/>100
WBCs (hpf) None = 0
Numerical ranges = 0-2/2-5/5-10/10-25/25-50/50-100/>100
Quality Assurance Errors
Preanalytical Patient misidentification
Wrong test ordered
Incorrect urine specimen type collected
Insufficient urine volume
Delayed transport of urine to the laboratory
Incorrect storage or preservation of urine
Analytical Sample misidentification
Erroneous instrument calibration
Reagent deterioration
Poor testing technique
Instrument malfunction
Interfering substances present
Misinterpretation of quality control data
Postanalytical Patient misidentification
Poor handwriting
Transcription error
Poor quality of instrument printer
Failure to send report
Failure to call critical values
Inability to identify interfering substances
TQM Based on a team concept involving personnel at all levels working together to achieve a
final outcome of customer satisfaction through implementation
CQI Improving patient outcomes by providing continual quality care in a constantly changing
health-care environment
PDCA Plan-Do-Check-Act
PDSA Plan-Do-Study-Act
OTHER BODY FLUIDS
Cerebrospinal Fluid
CSF 1st noted by Cotugno
Not an ultrafiltrate of plasma
Na+, Cl-, Mg2+: áCSF than in plasma
K+, Total Ca2+: âCSF than in plasma
3rd major body fluid
Production Filtration
Active transport secretion
Functions Supply nutrients à nervous tissue
Remove metabolic waste
Provide mechanical barrier
CSF glucose 60-70% of blood glucose
Brain 1,500g (Henry)
CSF Glucose
CSF Glucose NV = 60-70% of the plasma glucose concentration
A plasma glucose must also be run for comparison
Diagnostic significance:
-â values
-á CSF glucose values = result of á plasma glucose
Bulbourethral (Cowper’s) gland Secretes alkaline mucus à neutralize prostatic and vaginal acidity
Sertoli cells Serve as nurse cells for developing sperm cells
Inside the seminiferous tubules
Spermatogenesis Spermatogonia à 1’ Spermatocytes à 2’ Spermatocytes à Spermatids à Sperm
Round cells Either WBCs or spermatids
Seminal Fluid Composition
Seminal fluid (SV) 60-70%
Prostatic fluid 20-30%
Spermatozoa 5%
Bulbourethral gland 5%
Chemical Composition of Seminal Fluid
ACP For liquefaction
Zn3+ â in prostatic disease
Fructose Major nutrient of spermatozoa
K+, citric acid, ascorbic acid --
Proteolytic enzymes Liquefaction and coagulation
Spermine and Choline Inhibit growth of bacteria
Importance of Seminalysis To investigate the causes of infertility in marriages
To check the effectiveness of previous vasectomy
In medico-legal cases, where paternity is being disclamed on the basis of male sterility
Sexual abstinence 2-3 days and not >5 days
á abstinence = á volume, â motility
Methods of collection Important: 1st portion of ejaculate
1. Masturbation: best
2. Coitus interruptus (withdrawal method)
3. Common condom collection
-Condoms for sperm collection:
a. Silastic (Silicone rubber)
b. Polyurethane condoms: called the Male Factor Pak
4. Aspiration of semen from the vaginal vault after coitus
5. Specimen should be delivered in the lab w/in 1 hr (RT’)
Methods of preservation Specimen kept at 37’C awaiting analysis
For artificial insemination, it can be preserved in frozen state and stored at
-85’C (seminal banks)
Fresh specimen is clotted
Semen Analysis
Liquefaction time 30-60 mins
If not yet liquefied after 2 hrs, use α-chymotrypsin
Normal values Color = grayish white
Volume = 2-5 mL
Viscosity = pour in droplets
pH = 7.2-8.0
Sperm concentration = >20,000,000/mL
Sperm count = >40,000,000/ejaculate
Motility = >50% w/in 1 hr
Motility quality = >2.0 or a, b, c after 1 hr
WBCs = <1,000,000/mL
♫ >1,000,000/mL = inflammation
Volume â: incomplete collection/infertility
á: prolonged abstinence
Yellowish semen Prolonged abstinence
Medication
Urine contaminationi
á White turbidity Infection (á WBCs)
Red coloration (+) RBCs
Viscosity 0 (watery)
4 (gel-like)
pH Too basic = infection
Too acidic = á prostatic fluid
Sperm concentration Diluting fluid:
1. Cold H2O
2. Formalin
3. NaHCO3
4. 0.5% in chlorazene
5. 1% formalin in 3% trisodium citrate
1:20 = mechanical positive displacement pipette
Counting chamber 1. Neubauer counting chamber = diluted specimen
(WHO recommended)
2. Makler chamber = undiluted w/ heating processes
Purpose of Dilution To immobilize the sperm
Det. sperm conc. (Short-cut) 1. 5 RBC squares
# sperms counted x 1,000,000 = sperms in million/mL
2. 2 WBC squares
# sperms counted x 100,000 = sperms in million/mL
Sperm count Sperm concentration x volume of specimen
Motility quality (20/hpf) 4.0 (a) = Rapid motility
3.0 (b) = Slower speed, some lateral movement
2.0 (b) = Slow forward progression + lateral movement
1.0 (c) = No forward progression
0 (d) = No movement at all
CASA Computer-Associated Semen Analysis
-Sperm concentration
-Sperm velocity and trajectory
Sperm morphology At least 200 sperms evaluated
1. Routine criteria = >30% normal morphology
2. Kruger’s strict criteria = >14% normal morphology
-measure head, neck, tail using micrometer
Head morphology abnormalities Poor ovum penetration
Tail abnormality Poor motility
Sperm head Oval, approximately 5 µm x 3 µm wide
Tail 45 µm long
Midpiece Contains mitochondria
Connects head and tail
Acrosomal cap Ovum penetration
Covers approximately 2/3 of sperm nucleus and ½ of the head
Tapered head Varicocele
-Common cause of male sterility
-Hardening of veins that drains the testes
Stains Giemsa
Papanicolau = method of choice
Wright’s
Fructose test If sperm count is low
Rgt: Resorcinol
End-color: Orange-red
Specimens should be tested in 2 hrs or frozen
â neutral-α-glucosidase Epididymis disorder
Florence test Choline
(+) Dark brown rhombic crystals
Barbiero’s test Spermine
(+) Yellow leaf-like crystals
Spinbarkeit test Tenacity of mucus
Sim Huhner test Post-coital test
Test for the ability of sperm cells to penetrate the cervical mucosa
Bloom’s/Eosin-Nigrosin/ Sperm If N-sperm count but â motility
viability test Living sperm cells = bluish white
Dead = red
NV = 75% living sperms
(25% dead)
Decreased motility w/ clumping (+) Male antisperm antibodies
Blood testis barrier à disrupted
Clumps of sperm
Normal seminalysis w/ (+) Female antisperm antibodies
continued infertility
MAR Mixed agglutination reaction
Detect IgG antibodies
Immunobead test Detect IgG, IgA, IgM
Demonstrate area of the sperm the autoantibodies are affecting
Hamster egg penetration Sperm incubated w/ species non-specific hamster egg
Cervical mucus penetration Observed sperm penetration ability
Hypo-osmotic swelling Test for membrane integrity and viability of sperm
To determine whether semen is Microscopic exam for sperm cells
present Enhance w/ xylene
Examine à Phase microscope
Seminal glycoprotein p30: specific method
Aspermia No ejaculate
Azospermia Absence of sperm cells
Necrospermia Immotile/dead sperm cells
Oligospermia â sperm cells
Synovial Fluid
Synovial Latin: “Egg”
Fluid Diarthroses/joints
Arthrocentesis Method of collections
Synoviocytes Phagocytic cells
Secrete hyaluronic acid
Specimen collection Fluid à Syringe (heparin)
Micro: 3-5 mL in sterile tube, add 25 U heparin/mL fluid
Hema: 3-5 mL, add 25 U heparin/mL fluid
-Do not use crystalline EDTA but liquid EDTA may be accepted
Chem: 3-5 mL in red top and observe for clotting
NaF: glucose analysis
Normal Values Volume = <3.5 mL
Color = pale yellow
Clarity = clear
Viscosity = 4-6 cm long
Crystals = none present
Glucose = <10 mg/dL lower than the blood glucose
Lactate = <250 mg/dL
Total protein = <3 mg/dL
Uric acid = equal to blood value
Turbidity á WBCs
Milky (+) Crystals
Rope’s/Mucin clot test Hyaluronate polymerization test
2-5% acetic acid
Grading:
-Good = solid clot
-Fair = soft clot
-Low = friable clot
-Poor = no clot at all
Cells in Synovial Fluid
WBC Total WBC count: most frequently tested
STAT, otherwise, refrigerate
Neubauer counting chamber
Clear fluids = counted undiluted
Diluting fluid = NSS
If necessary to lyse RBCs:
-Hypotonic saline (0.3%)
-Saline w/ saponin
Do not use WBC diluting fluid
Differential count 65% = Monocytes & Macrophages
<25% = Neutrophils
<15% = Lymphocytes
Very viscous specimen Add hyaluronidase à 0.5 mL of fluid or
1 drop of 0.05% hyaluronidase in PO4 buffer/mL of fluid
37’C for 5 mins
LE cell Neutrophil
Reiter cells Vacuolated macrophage w/ ingested neutrophils
Ragocyte Neutrophil w/ dark cytoplasmic granules
Cartilage cells Large multinucleated cells
Rice bodies Macroscopically resembles polished rice
Ground pepper appearance of Ochronotic shards = debris
synovial fluid Metal and plastic joint prosthetic
Hemosiderin Pigmented villonodular synovitis
Pericardial Fluid
Pericardiocentesis Method of collection
Normal volume 10-50 mL
Mesothelial cells Produces pericardial fluid
Function Reduces friction
Normal appearance Clear/pale yellow
Grossly bloody Accidental cardiac puncture
Misuse of anticoagulant medicatios
Milky Chylous/pseudochylous effusions
Bacterial endocarditis >1000 WBCs/µL (á% neutrophils)
Peritoneal Fluid (Ascites)
Paracentesis Method of collection
Peritoneal lavage Sensitive test for the detection of intra-abdominal bleeding
WBC <500/µL Normal
RBC >100,000/µL Blunt trauma cases
Serum Ascites Albumin Recommended to differentiate transudates from exudates
Gradient (SAAG) a. >1.1 = Transudate
b. <1.1 = Exudate
Psammoma bodies Contains concentric striations of collagen materials
Ovarian and thyroid malignancies
(+) CA 125 Tumor of ovaries, fallopian tubes or endometrium
(-) CEA
Glucose â TB peritonitis
ALP Intestinal perforation
AMS Pancreatitis
GI perforation
BUN Ruptured bladder or accidental puncture of the bladder
Creatinine
Sweat Test
Cystic fibrosis (mucoviscidosis) Autosomal recessive
Pancreatic insufficiency
Respiratory distress
Intestinal obstruction
-Bulky offensive greasy stools (butter-like)
Gibson and Cooke Pilocarpine á Sweat Cl- and Na+ = >70 mEq/L
Iontophoresis Borderline = 40 mEq/L
Pilocarpine w/ mild current = stimulates sweat glands
Sodium FEP, IEE
Chloride Manual or automated titration
Amniotic Fluid
During 1st trimester 35 mL: derived from maternal circulation/plasma
After 1st trimester Fetal urine
rd
3 trimester Peak: 1L amniotic fluid
Oligohydramnios â amniotic fluid
Urinary tract deformities
Membrane leakage
á fetal swallow of urine
Hydramnios/ á amniotic fluid
Polyhydramnios â fetal swallow of urine
Neural tube defects
Amniocentesis Method of collection
2nd trimester: assess genetic defects
3rd trimester: FLM or HDN
Fern test Specimen: Vaginal fluid
Glass slide à air dry
(+) Fern-like crystals
(+) Amniotic fluid
Normal appearance Clear
Green Meconium
Yellow Bilirubin (HDN)
Bloody/red Trauma, abdominal trauma, intra-amniotic hemorrhage
Dark-brown/ Fetal death
Reddish-brown
Assessment of HDN á OD at 450 nm
OD reading at a Liley graph
1. Zone 1: observe fetus for stress
2. Zone 2: moderate disease
3. Zone 3: severe disease