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PREPARED BY: Ma’am Jennifer Gaytano-Bautista

PART IV. OTHER BODY FLUIDS Tube # Section Storage
1 Chemistry or Frozen
serology section
CEREBROSPINAL FLUID 2 Microbiology Room
temperature
CSF 3 Hematology Refrigerated

➔ Considered as the 3rd major body fluid APPEARANCE

➔ 1st recognized by Catugno in 1764
➔ Does not resemble an ultrafiltrate of plasma Crystal Clear Normal
➔ CSF is derived from ultrafiltration and secretion Cloudy, Milky, Turbid ↑ Protein, Lipid, WBC
of blood through choroid plexus Bloody Intracranial hemorrhage,
traumatic tap
Functions Xanthochromic (Pink, Hemoglobin, bilirubin,
Orange, Yellow) carotene, ↑ protein,
➔ Supply nutrients to nervous tissue melanin
➔ Removes metabolic waste ➔ Pink – oxyhemoglobin
➔ Orange – heavy hemolysis
➔ Cushion the brain & spinal cord against trauma
➔ Yellow – conversion of oxyhemoglobin into
CSF Volume unconjugated bilirubin
Clotted Clotting factors
➔ Adults: 90-150 mL introduced by traumatic
type
➔ Neonates: 10-60 mL
Pellicle Tubercular meningitis
➔ Production of CSF: 20 mL/hr approx. 480 mL/d seen after overnight of
➔ In Henry’s approx. 500 mL/d refrigeration
Oily Radiographic contrast
Formation & Physiology
media
➔ Meninges – lines the brain and the spinal cord TRAUMATIC TAP VS. INTRACRANIAL
3 layers: HEMORRHAGE
➔ Dura mater: hard matter; outer most layer of
the meninges; it lines the skull and vertebral Traumatic Tap Intracranial Hemorrhage
canal Uneven distribution of Even distribution of blood
➔ Arachnoid matter – middle layer; spider blood
like/web like structure Clot formation No clot formation
➔ Pia mater – inner layer; lines the surface of Supernatant: not Supernatant:
the brain and spinal cord xantochromic xantochromic
➔ Subarachnoid space – space between the Erythrophagocytosis -
arachnoid matter and pia mater; this is where macrophage containing
the CSF flows RBCs/hemosiderin
➔ Choroid plexus - Responsible for the granules
(+) D-dimer – indication of
production of CSF
fibrin degradation product
➔ Blood-Brain-Barrier (BBB) – tight fitting
endothelial cells Cell Count
➔ Produces 20 mL/hr of fluid
➔ Most routinely performed WBC count
Specimen Collection
➔ Should be performed ASAP
➔ RBC: disintegrate within 1 hour
➔ Procedure: lumbar tap/lumbar puncture: 3rd, 4th,
➔ ↓ by 40% in WBC: after 2 hours
5th lumbar vertebrae
➔ Pleocytosis - ↑ number of cells in CSF
➔ Before aspirating: opening pressure is
measured first Methods
➔ Ideally: test is performed on STAT basis
➔ Position: lying or sitting position ➔ Manual – Neubauer counting chamber
➔ Automated: automated cell counters
CSF Specimen Collection Tubes
➔ QC: results of manual count should agree with
automated count by ± 25%
➔ 3 sterile tubes: numbered in order they are
withdrawn Normal Values
➔ If possible: 4th tube may be withdrawn for
microbiology section ➔ Adult: 0-5 cells/uL → predominant cells:
LYMPHOCYTES

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➔ Neonates: 0-30 cells/uL → predominant cells: ➔ Level declines rapidly when treatment is
MONOCYTES successful

CSF Electrophoresis CSF Glutamine

➔ Primary Purpose: detection of oligoclonal ➔ Preferred over direct measurement of CSF

bands ammonia
➔ Can determine if a fluid is indeed CSF ➔ Ammonia is volatile
➔ Normal values: 8-18 mg/dL
Oligoclonal Bands ➔ Elevated levels: associated with liver disorder
that result in ↑ blood and CSF ammonia
➔ Indicates immunoglobulin production ➔ Frequently requested procedure for patients
➔ Bands are located in the GAMMA region of the with coma of unknown origin
protein electrophoresis ➔ ↑ in approximately 75% of children with Reye’s
➔ Interpretation: syndrome
 Oligoclonal bands in CSF ONLY: ➔ Glutamine produced from ammonia & α-
▪ Multiple sclerosis ketoglutarate by the brain cells
▪ Other: neurologic disorder,  This process serves to remove
encephalitis, neurosyphilis, ammonia from the CNS
Guillain-Barre syndrome
 Oligoclonal Bands in CSF and Microbiology Test
SERUM
▪ HIV infection ➔ Identification of causative agent in Meningitis
 Oligoclonal Bands in SERUM ONLY ➔ Microorganism must be recovered by growing it
▪ Leukemia, Lymphoma, Viral on the appropriate culture medium
infections ➔ 24 hours in cases of bacterial meningitis
▪ May produce CSF banding ➔ 6 weeks for tubercular meningitis
because of BBB leakage or ➔ Specimen concentration: CSF centrifuged @
blood contamination during 1500 g for 15 minutes → sediment used or slides
lumbar tap & culture
➔ CSF culture: confirmatory test rather than
Myelin Basic Protein diagnostic test
➔ Microbiology test for a preliminary diagnosis
➔ Indicates recent destruction of the myelin sheath  Gram stain
that protects the axon of the neurons  Acid fast stain
(demyelination)  India ink preparation
➔ Measurement can be used to monitor the course  Latex agglutination test
of multiple sclerosis
MAJOR LABORATORY RESULTS FOR THE
CSF Glucose
DIFFERENTIAL DIAGNOSIS OF MENINGITIS
➔ Value: 60-70% of plasma glucose Bacterial Viral Tubercular Fungal
➔ Example: Neutrophil Lymphoc Lymphocyt Lymphocytes,
 Plasma glucose: 100 mg/dL
Predomina

ytes es, monocytes,

nt Cell

 CSF glucose: 65 mg/dL monocytes eosinophil

➔ Blood glucose: should be drawn 2 hours prior
to lumbar tap (CSF)
➔ ↑ levels: always a result of plasma elevations of
Marked Moderate Moderate to marked
glucose
↑ Protein

➔ ↓ levels: caused by alteration in transport in

BBB and ↑ used by brain cells

Lactate
Markedly ↓ Normal ↓ Normal to ↓
Glucose

➔ Destruction of tissue within the CNS due to

hypoxia causes the production of ↑ CSF lactate
➔ Aid in the diagnosis and management of
meningitis cases
➔ Used to monitor severe head injuries >35 mg/dL >25 mg/dL
➔ Falsely elevated: obtained from xantochromic or
Lactate

hemolyzed samples
➔ Elevation is consistent with: bacterial,
tubercular, and fungal meningitis

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Testes ➔ Seminiferous tubules –
production of
Gram Stain spermatozoa
Epididymis ➔ Storage and maturation
➔ Routinely performed on CSF from all suspected of sperm
cases of meningitis Seminal ➔ Produces most of the
➔ Values lies in the detection of bacterial and Vesicles fluid present in the semen
fungal organism (60%-70%)
➔ Use of cytocentrifuge: provides highly ➔ Fructose – used by
concentrated specimens sperm for energy &
motility
COMMON CAUSES OF BACTERIAL MENINGITIS ➔ Flavin – responsible for
gray appearance of
Infants ➔ S. agalactiae sperm
➔ E. coli K1 strain, L. Prostate Gland ➔ Aids in propelling the
monocytogenes sperm through the
Adolescents/Adults ➔ H. influenzae type B, urethra by contractions
S. pneumoniae during ejaculation
➔ S. pneumoniae Bulbourethral ➔ Contributes about 5% of
Elderly ➔ N. meningitides, L. Gland the fluid volume in the
monocytogenes form of thick alkaline
➔ C. neoformans mucus (helps neutralize
vaginal acidity)
Cryptococcus neoformans
SEMEN ANALYSIS
➔ Cryptococcal meningitis (one the frequently
occurring complications of AIDS) ➔ Sexual abstinence – at least 2 days but not
➔ Laboratory findings: India ink: thick more than 7 days
encapsulated organism  Prolong abstinence – higher volume
➔ Gram stain: starburst pattern (may be seen and decreased motility
more often than a positive India ink ➔ Fertility testing – according to WHO: 2 or 3
➔ Associated with ↑ eosinophils sample be collected not les than 7 days or more
than 3 weeks apart with 2 abnormal samples
Limulus Lysate Test considered as significant
➔ Specimen/Semen – if possible, should be
➔ Diagnosis of meningitis caused by gram (-) collected in the room provided by the laboratory
bacteria  Should be analyzed within 1hr
➔ Limulus amoebacyte reacts with bacterial  Semen collected outside the laboratory
endotoxin of gram (-) bacteria should be kept in room temp – 20-24°C
➔ Reagent: from blood cells of horseshoe crab and should be delivered in the
(Limulus polyphemus) laboratory 1hr before the collection
➔ (+) result: coagulation within 1 hour of  37°C temperature if the semen will not
incubation @37°C be processed immediately
 Should be collected by masturbation
Serologic Testing  Non-lubricant condom and
rubber/polyurethane condom can be
➔ Detection of neurosyphilis
used in collection
➔ Venereal Disease Research Laboratories
(VDRL) PARAMETERS
➔ Procedure recommended by the CDC to
diagnose neurosyphilis APPEARANCE
➔ Fluorescent Treponemal Antibody-Adsorption
(FTA-ABS) Test - more sensitive than VDRL; ➔ Normal semen – gray-white color, appears
prevent contamination with blood because the translucent, has characteristic of musty odor
FTA-ABS remains positive in the serum of ➔ Low sperm concentration: appears almost
treated cases of syphilis clear
➔ Increase white turbidity: indicates presence of
SEMINAL FLUID WBCs and infection within the reproductive tract
➔ LE reagent strip: useful in differentiating
Semen presence of WBC compare to immature sperm
(spermatids)
➔ Composed of 4 fractions that are contributed by: ➔ Red coloration: presence of RBC

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➔ Yellow coloration: urine contamination – ➔ Sperm count: sperm concentration x sperm
retrograde ejaculation (significant finding), volume
prolonged abstinence and medication ➔ Normal sperm count: >40 million per ejaculate

Motility

Liquefaction ➔ Sperm should be capable of forward,

progressive motility
➔ Fresh semen specimen is clotted within 30-60 ➔ Should be assessed using a well-mixed,
mins after collection liquefied specimen within 1hr of specimen
➔ Failure of liquefaction w/in 60 mins.: caused collection
by deficiency in prostatic enzymes and should
be reported Morphology
➔ Prostate gland: produces 20-30% acidic fluid
➔ Milky acidic fluid components: acid ➔ Head – oval shape approx. 5um long and 3um
phosphatase, citric acid, zinc, proteolytic wide
enzyme  Acrosomal cap – enzyme containing
 Responsible for coagulation and critical to ovum penetration
liquefaction of semen (encompasses half of the head and
➔ If after 2 hours the specimen has not liquefied, cover approx. two thirds of the sperm
an equal volume of physiologic Dulbecco’s nucleus)
phosphate-buffered saline of proteolytic ➔ Neckpiece – attaches the head to the tail and
enzymes such as alpha-chymotrypsin or midpiece
bromelain may be added to induce liquefaction ➔ Midpiece – approx. 7um long, thickest part of
and allow the rest of the analysis to be the tail (surrounded by the mitochondrial sheath
performed that produces energy required by tail for motility)
➔ Tail – 45 um long
Volume
Criteria
➔ Normal semen volume: 2-5 mL
➔ Increased volume: prolonged abstinence ➔ Strict criteria – normal form >14%
➔ Decreased volume: frequently associated with ➔ WHO Criteria - >30%
infertility ➔ Round cells - <1.0 M/mL
➔ Incomplete specimen collection must be ➔ Air-dried smear – used to test for morphology
considered ➔ Stains used
 Wright’s stain
Viscosity  Giemsa
 Shorr
➔ Refers to the consistency of the fluid and may  Papanicolaou stain
be related to specimen liquefaction ➔ Evaluate 300 sperm - >14% normal form should
➔ Incomplete liquefied specimen: clumped and be obtained
highly viscous
➔ Normal: easily drawn into a pipette and form Abnormal Heads
small discrete droplets, do not appear clumped
or stringy ➔ Double head
➔ Droplets that form longer than 2cm: highly ➔ Giant/amorphous pinheads
viscous, abnormal ➔ Tapered
➔ Constricted
pH
Tails
➔ Normal pH: 7.2-8.0
➔ Increased pH: indicates infection within the ➔ Doubled
reproductive tract ➔ Coiled
➔ Decreased pH: increased prostatic fluid, ➔ Bent
ejaculatory duct obstruction, poorly developed
seminal vesicles ADDITIONAL TESTS

Sperm Concentration Sperm Vitality

➔ Normal sperm concentration: >.20-250 mL ➔ Normal sperm concentration is tested

sperm/mL ➔ ↓ SV – normal sperm concentration, markedly
➔ Borderline sperm concentration: 10-20 decreased motility
million per sperm/mL

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➔ Eosin Nigrosin Stain – 100 sperm is evaluated; ▪ T. vaginalis
dead cells are counted; phase contrast  Yellow, opaque cervical discharge:
microscope is used ▪ C. trachomatis
 NSV - >50 living sperm cells %; will
remain bluish white DIAGNOSTIC TEST/S PARAMETERS
 Dead sperm cell – stains red against
purple background pH

➔ Normal vaginal flora, bacteria, lactobacilli:

VAGINAL FLUID SECRETIONS
produces lactic acid (end product of glycogen
metabolism), provides an acidic vaginal
Vaginal Secretions
environment with pH between 3.8-4.5H2
➔ H2O2 – keeps the vaginal acidic and provides
➔ Examined in order to diagnose infection,
protection against infections
complication of pregnancy, forensic testing in
➔ >4.5 pH – bacterial vaginosis, Trichomoniasis,
sexual assault patients
desquamative inflammatory vaginitis, atrophic
Specimen Collection & Handling vaginitis
➔ 3.8-4.5 – Candidiasis
➔ Speculum – moistened with warm water is used ➔ Estrogen – production is necessary to preserve
to visualize vaginal fornices an acidic vaginal environment
➔ Collected by: swabbing the vaginal walls and
vaginal pool to collect the epithelial cells along Microscopic Procedure
with vaginal secretions
➔ Initial Screening Test: saline wet mounts &
➔ Sterile, polyester-tipped swab on plastic
KOH
shafts or swabs – specifically designated by
 KOH – used to identify yeast cells;
manufacturer should be used
lyses RBC
➔ Cotton swabs – toxic to N. gonorrhea
 Amine – whiff test
➔ Wood in wooden shafts – toxic to Chlamydia
➔ Confirmatory Test (Yeast Bacterial
trachomatis
Vaginosis) – gram stain
➔ Calcium alginate – can inactivate herps
simplex virus (HSV) for viral cultures
➔ Specimen must be kept in: QUANTITATION SCHEME FOR MACROSCOPIC
 Room temperature to:
ANALYSIS
▪ Preserve motility of T.
vaginalis Rare <10 organisms or cells /slide
▪ Recovery of N. gonorrhea 1+ <1 organisms or cell /HPF
 Refrigerate 2+ 1-5 organisms or cells /HPF
▪ For C. trachomatis and HSV 3+ 6-30 organisms or cells /HPF
to prevent overgrowth of 4+ >30 organisms or cells /HPF
normal flora
WET MOUNT PREPARATION
Color & Appearance
Squamous EC
➔ Normal vaginal fluid appears white with
flocculent discharge ➔ Exhibit a polygonal “flagstone” appearance
➔ Microscopically:
Clue Cells
 Normal vaginal flora: predominance of
large, rod-shaped, gram + lactobacilli
➔ Squamous epithelial cells with attached
 Squamous epithelial cells
coccobacillus
 WBC and RBS may be present if the
➔ Granular, irregular appearance
patient is menstruating
➔ “shaggy”
➔ Abnormal vaginal secretions
➔ Diagnostic for bacterial vaginosis (G. vaginalis)
 Homogenous white to gray
discharge White Blood Cells
▪ Often seen in bacterial
vaginosis ➔ Are present in rare to scanty numbers in vaginal
 White “cottage-cheese” like secretions
discharge ➔ >3+ WBCS: vaginal candidiasis, atrophic
▪ Particular for Candida vaginitis, or infections with Trichomonas,
infections Chlamydia, N. gonorrhea, herpes simplex
 Yellow-green frothy, adherent ➔ Present due to menstruation or desquamative
discharge inflammatory process

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➔ Confused with yeast cells, can be distinguished NUGENT’S GRAM STAIN CRITERIA TO DIAGNOSE
by KOH → RBC will lyse BACTERIAL VAGINOSIS
Red Blood Cells Lactobacillus Gardnerella Curved Points
& gram-
➔ Present due to menstruation or desquamative Bacteroides variable
inflammatory process spp. rods
➔ Confused with yeast cells, can be distinguished Morphocytes
by KOH → RBC will lyse 4+ 0 0 0
3+ 1+ 1+ or 2+ 1
Parabasal Cells 2+ 2+ 3+ or 4+ 1
1+ 3+ 3
➔ Round to oval shaped with marked basophilic 0 4+ 4
granulation or amorphic basophilic structures NOTE: points are added according to morphocytes seen.
(“blue blobs”) in surrounding cytoplasm Add the points for all 3 columns for a final sum. A score of
➔ Increased in desquamative inflammatory 7 or higher indicates BACTERIAL VAGINOSIS
vaginitis accompanied by large numbered of
WBCs KOH Preparation and Amine (Whiff) Test

Basal Cells ➔ Performed by placing a drop of saline specimen

prepared from the collection swab onto a
➔ Round cells which can be distinguished from properly labeled clean slide and adding one drop
WBCs (multilobed nucleus) of 10% KOH
➔ Large number of WBCs: indicative of ➔ Check for “fishy” amine odor
desquamative inflammatory vaginitis ➔ Positive – presence of fishy odor; negative –
absence of fishy odor
Trichomonas vaginalis
Gram Staining
➔ Atrial flagellated protozoan that can cause
vaginal inflammation and infection in women ➔ Considered as the gold standard in identifying
➔ Exhibits “jerky” motion observed in wet mount the causative organisms for bacterial vaginosis
➔ Dead Trichomonas tend to appear oval and
slightly larger than a WBC Culture

Yeast Cells ➔ Considered gold standard for detecting yeast &

Trichomonas – time consuming & requires up to
➔ Candida albicans and non-Candida spp. causes 2 days of result)
most fungal infections but an occasional yeast ➔ Diamond’s Medium – special media required
cell is considered as part of normal flora for T. vaginalis
➔ Appears in wet mount as both budding yeast ➔ Culture for G. vaginalis – not a diagnostic
cells (blastophores) or has hyphae, which are procedure for bacterial vaginosis – part of
long filaments and from a mycelium normal flora in 50% of healthy women

Bacteria AMNIOTIC FLUID

➔ Vagina – non-sterile environment with complex
PHYSIOLOGY
endogenous bacterial flora that varies with age
& hormonal status of patient ➔ Amnion – membranous sac that surrounds the
➔ Lactobacillus spp. – largest portion of bacterial fetus
flora
 Appearance: large, gram +, non-motile PRIMARY FUNCTION OF AMNIOTIC FLUID
rods
 Produces lactic acid, maintains the ➔ Provide protective cushion for the fetus
vaginal pH @ 3.8-4.5 ➔ Allow fetal movement
 H2O2 produced by lactobacillus spp. ➔ Stabilize the temperature to protect the fetus
helps to suppress growth of other from extreme temperature changes
organisms ➔ Permit proper lung development
➔ Mobiluncus spp. – thin, curve, gram-negative
motile rods VOLUME
➔ Gardnerella vaginalis/Bacteroides spp. – short, AF volume is regulated by a balance between:
gram-negative coccobacilli
1. Production of Fetal urine & lung fluid

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2. Absorption from fetal swallowing & ▪ + Screen for AMNIOTIC
intramembranous flow FLUID: presence of “FERN-
a. 3rd Trimester – approximately 800- LIKE” crystals (due to protein
1200 mL (gradually decrease prior to & sodium chloride)
delivery)
b. Polyhydramnois – Amniotic Fluid Amniocentesis
>1200 mL
i. Due to: failure of the fetal ➔ AF is obtained by needle aspiration into the
lung to begin swallowing amniotic sac
ii. Secondary associated with: ➔ Maximum of 30mL collected in sterile syringe
fetal structural anomalies, ➔ 1st 2-3mL collected → DISCARDED
cardiac arrhythmias, ➔ Safe if performed after 14th week gestation
congenital infections, Procedure
chromosomal abnormalities
c. Oligohydramnois – Amniotic Fluid < 1. Transabdominal Amniocentesis: most
800 mL frequently performed
i. Due to: ↑ fetal lung 2. Vaginal Amniocentesis: great risk of infection
swallowing, urinary tract
deformities, membrane Specimen Handling & Storage:
leakage
➔ Bilirubin Testing
COMPOSITION OF AMNIOTIC FLUID  Protected from light
 Place in amber colored tubes or black
1st Trimester plastic cover for container
➔ Fluid for Chemical Testing
➔ Volume of approximately 35mL
 Separated from cellular element and
➔ Composition similar to maternal plasma
debris
➔ Contains small amount of sloughed fetal cells
➔ Cytogenetic Studies
 Basis for Cytogenetic Analysis
 Stored at RT or Body Temperature
3rd Trimester (370C)
➔ FLM (Fetal Lung Maturity)
➔ Volume reaches a peak of 1L – gradually  Low speed centrifugation not >5
decreases prior to delivery minutes
➔ Major Volume Contributor in Fetal Urine  Filtration recommended prior to testing
o ↑ Creatinine, Urea, Uric Acid  Delivered in ICE
o >2mg/dL Creatinine = Fetus >36  Refrigerated prior to testing (Tested
weeks within 72 hours)
o AF Creatinine does not exceed
AF Color & Appearance
3.5mg/dL & Urea 30mg/dL
o ↓ Glucose & Protein
➔ Normal AF: Colorless
 Transparency: slight to moderate
DIFFERENTIATING MATERNAL URINE FROM
turbidity (from cellular debris,
AMNIOTIC FLUID particularly in later stages of fetal
Maternal Urine Amniotic Fluid development)
Creatinine High (up to Lower (don’t
Color Significance
10mg/dL) exceed
Colorless Normal
3.5mg/dL)
Blood-streaked Traumatic Tap, abdominal
Urea High (up to Lower (don’t
trauma, intra-amniotic
300mg/dL) exceed
hemorrhage
30mg/dL)
Yellow HDN (Bilirubin)
Glucose & Negative (Normally) Present
Dark Green Meconium (NB 1st bowel
Protein
movement)
Fern Test Negative Positive
Dark red-brown Fetal death

➔ Fern Test – used to evaluate premature rupture TESTS FOR FETAL DISTRESS
of the membranes.
 Vaginal Fluid specimen is spread on
Hemolytic Disease of the Newborn (HDN)
the glass slide
➔ Oldest routinely performed laboratory test on
 Allow to completely dry at RT
amniotic fluid evaluates the severity of the fetal
 Observed microscopically:
anemia produced by HDN.

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➔ Rh-Negative Mother ↔ Rh-Positive Newborn ➔ Sphingomyelin: lipid that is produced at a
 Initial exposure to foreign red cell constant rate after about 26th week of gestation
antigen occurs during: (STIMULATES
the mother to produce antibodies L/S Ratio
against the antigen) Prior to 35 weeks’ Usually, <1.6
▪ Gestation gestation
After 35 weeks’ 2.0 or higher
▪ Delivery of the placenta
gestation
▪ Previous pregnancy (when Therefore: when L/S Ratio reaches 2.0 → PRETERM
fetal RBCs enter the maternal delivery is usually considered to be relatively a
circulation SAFE procedure
➔ Presence of (red blood cell degradation ➔ Falsely Elevated L/S Ratio: AF contaminated
product): Unconjugated Bilirubin (Amniotic with blood or meconium
Fluid) → due to destruction of fetal red blood
cells Phosphatidyl Glycerol

Neural Tube Defects (NTD) ➔ Lung surface lipid

➔ Can be detected after 35 weeks’ gestation
➔ Most common birth defects in the US ➔ Production of PG normally – parallel – with –
➔ Can be detected by: Maternal Serum Alpha- LECITHIN
Fetoprotein (MSAFP), High Resolution  Production is DELAYED: in cases of
Ultrasound & Amniocentesis Maternal Diabetes
➔ ↑ AFP (both maternal circulation & AF):
Indicative of NTD (e.g., Anencephaly & Spina Foam Stability Index
Bifida)
➔ AFP: major protein produced by the fetal liver ➔ “Foam” or “Shake” Test
during early gestation (prior to 18 weeks)  Mechanical Screening Test
➔ Amniotic Acetylcholinesterase (AChE) – more  Measure the INDIVIDUAL LUNG-
specific than AFP SURFACE lipid concentrations
 Not to be performed in bloody
Procedure:
specimen → Blood contains AChE
1.Amniotic Fluid + 95% Ethanol
TESTS FOR FETAL LUNG MATURITY 2.Shake for 15 seconds
Fetal Lung Maturity 3.Allowed to sit undisturbed for 15 minutes
4.Surface of the fluid is observed for: PRESENCE
➔ Respiratory Distress Syndrome (RDS): OF CONTINUOS LINE OF BUBBLES AROUND
 Most frequent complication of early THE OUTSIDE EDGE.
delivery a. Presence of the Bubbles: Indicates a
 7th most common cause of morbidity & sufficient amount of phospholipids
mortality in premature infant ➔ Falsely Mature Index Result: AF contaminated
 Cause by insufficiency of Lung with blood or meconium
Surfactant production & structural
Lamellar Bodies
immaturity of the fetal lungs
➔ Surfactant: ➔ Densely packed layers of phospholipids that
 Normally appears in mature lungs and represent a storage form of pulmonary
allows the alveoli (air sacs of the lung) surfactant
to remain open throughout the normal ➔ Secreted by the type II pneumocytes of the fetal
cycle of inhalation and exhalation lung at about 24 weeks of gestation
 Keeps the alveoli from collapsing by ➔ Absorbed into the alveolar spaces to provide
decreasing surface tension and allows surfactant
them to inflate with air more easily ➔ Enters the AF at about 26th week gestation
 ↓ Surfactant = Collapsed of Alveoli = ➔ Increase in concentration from 50,000–
RDS 200,000/mL by the end of 3rd Trimester
Lecithin-Sphingomyelin Ratio ➔ Fetal Lung Mature = ↑ Lamellar Body Production
= ↑ Amniotic Fluid Phospholipids & L/S Ratio
➔ L/S Ratio: reference method for FLM
➔ Lecithin: primary component of the surfactants
(phospholipids, neutral lipids & proteins)
 Produced at a relatively low & constant
rate until the 35th week of gestation

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