unit 6 Body Fluids – Quick Reference Notes (Last-Minute)
Cerebrospinal Fluid (CSF)
- Definition and role: clear, colorless, sterile fluid circulating in brain ventricles, subarachnoid spaces, and central canal; cushions CNS and provides nutrient medium; not an ultrafiltrate of plasma due to blood–brain barrier
- Volume and composition: adults have 90\text{--}150\,\text{mL}; contains ~99% water; no RBCs; low protein and lipids compared with serum; CSF protein ~35\,\text{mg/dL}; glucose ~60\,\text{mg/dL}; pH ~7.33; osmolarity ~295\,\text{mOsm/L}
- Four main functions: mechanical buffering, intracranial volume regulation, CNS nutrition, excretion of CNS metabolites
- Indications for collection: meningitis diagnosis, cerebral hemorrhage, neurological disease, suspected malignancy, administration of drugs/contrast
- Sample handling (CSF):
- Collect via lumbar puncture under sterile conditions; deliver STAT to lab immediately
- Typical collection: 3–4 sterile tubes (~1 mL each), labeled 1–4
- Tube roles:
- Tube 1: chemistry (e.g., glucose, protein, xanthochromia)
- Tube 2: microbiology (Gram stain, culture)
- Tube 3: hematology (total cell counts, differential)
- Tube 4: cytology
- Storage and retention:
- Some tests refrigerated or frozen; microbiology tubes at room temp
- Follow SOPs for CSF storage; retention varies by department
- Analysis goals:
- Diagnose CNS disorders: meningitis, encephalitis, hemorrhage, tumors, MS, syphilis
- Biomarkers: e.g., amyloid beta, tau (Alzheimer risk) as applicable
- Visual and microscopic assessment:
- Record colour, clarity, volume for each tube
- Traumatic tap vs hemorrhage: RBC distribution across tubes differs; traumatic tap shows decreasing RBCs from Tube 1 to Tube 4; if uniform RBCs across all tubes, suggests genuine hemorrhage
- Common CSF analyses:
- Routine chemistry: protein, glucose, lactate
- Microbiology: Gram stain, culture; viral testing as indicated
- Cytology and morphology: cytocentrifugation preferred for cell analysis
- Hematology: total WBC count and differential; CSF usually 0–8×109/L in adults; higher counts with infection or inflammatory conditions
- Cytocentrifugation (CSF):
- Use specialized cytocentrifuge; slow spin improves cell yield and morphology
- Procedure: mix ~0.5\,\text{mL} CSF with a few drops of bovine serum albumin; centrifuge slowly; transfer cells to slide; stain (e.g., Wright)
- Smear preparation when cytocentrifuge unavailable: regular centrifuge; sediment onto slides; cells may be distorted
- Routine CSF chemistry and microbiology highlights:
- Protein increased with meningitis, hemorrhage, MS; glucose reduced in bacterial infection; levels should be measured promptly for glucose
- Lactate used in meningitis evaluation
- Biomarkers for CNS diseases and microbiology tests (Gram stain, culture, TB tests, VDRL for syphilis)
Serous Fluids
- What they are: pleural (lungs), pericardial (heart), and peritoneal/ascitic (abdomen); normal appearance pale yellow; small volumes (pleural 1–10 mL typically)
- Effusions: accumulation of serous fluid; differentiate transudates vs exudates
- Transudates vs Exudates (key definitions):
- Transudates: systemic factors altering pressure/filtration; e.g., CHF, liver dysfunction, nephrotic syndrome
- Exudates: local inflammation or infection affecting membranes; e.g., infections, malignancy, inflammatory diseases
- Differentiation criteria (fluid vs serum):
- Fluid:serum protein ratio < 0.5 (transudate) or > 0.5 (exudate)
- Fluid:serum LDH ratio < 0.6 (transudate) or > 0.6 (exudate)
- WBC counts: < 1000/µL (transudate) vs > 1000/µL (exudate)
- Spontaneous clotting: unlikely in transudates; may occur in exudates
- Pleural fluid cholesterol ratio and bilirubin ratio help differentiation
- Serum-ascites albumin gradient (SAAG): > 1.1 indicates transudate; < 1.1 indicates exudate
- Pleural fluid testing significance:
- Glucose: decreased in rheumatoid/inflammatory or purulent infection
- Lactate: elevated with bacterial infection
- Triglyceride: elevated in chylous effusions
- ADA, Amylase: used for TB and other conditions or pancreatitis, respectively
- Pericardial fluid: normal volume ~25 ext{--}50\,\text{mL}; testing includes Gram stain/culture, ADA for TB, cytology for malignant cells
- Peritoneal fluid: normally < 100\,\text{mL} in cavity; turbid fluids indicate infection; TB may show lymphocyte predominance
- Collection guidelines for serous fluids:
- Typically 3 tubes: EDTA for cells and morphology; heparin for chemistry; sterile for Gram stain/culture
- Optional additional tubes for cytology
Synovial Fluid
- What it is: fluid within joints; viscous ultrafiltrate of plasma enriched with hyaluronate; hyaluronic acid gives normal viscosity
- Normal characteristics: straw-colored, viscous; WBC < 200\,/\text{mL}; mostly mononuclear cells; sterile; crystals absent
- Indications for arthrocentesis: diagnose joint disease (crystal-induced, degenerative, inflammatory, infectious)
- Classification of joint fluid:
- Noninflammatory (e.g., osteoarthritis, traumatic arthritis)
- Inflammatory (e.g., RA, lupus arthritis)
- Infectious (bacterial infection)
- Crystal-induced (e.g., gout)
- Hemorrhagic (bleeding in joint, tumor, trauma)
- Collection and processing:
- Three tubes: microbiology (sterile), liquid EDTA or heparin (microscopy), plain tube for clotting and chemistry/cytology
- Routine examination:
- Gross appearance, viscosity, WBC count, differential, and morphologic examination (cytocentrifuged slides preferred)
- Microscopy for crystals
- Microbiological testing: Gram stain and culture (bacteria, mycobacteria, fungi)
- Chemistry tests: glucose (lower in infected joints), protein (increased in RA, gout, infections)
- Other tests: LDH, uric acid, lactate; immunologic tests (RF, ANA)
Seminal Fluid
- Context: semen analysis for fertility, forensic purposes, vasectomy assessment, ART suitability
- Specimen handling:
- Fresh specimen; ideally delivered within 30\,\text{minutes}; kept at 37^{\circ}\text{C}; examined within 2\ hours
- Abstinence period: 3–5 days; avoid condom use prior to collection
- Transport: keep warm if collected at home; prompt delivery to lab
- Sperm count reference: 20\text{--}160\ times 10^6/\text{mL}
- Macroscopic analysis: time to complete liquefaction, appearance, volume, viscosity, pH
- Wet-mounted analysis: approximate sperm count and motility
- Other assessments: viability, agglutination, counting-chamber counts, morphology on stained slides
- Indications: fertility assessment, forensic purposes, vasectomy evaluation, ART suitability
Amniotic Fluid
- Definition: nourishing, protective fetal liquid; clear, yellow, primarily water with proteins, carbohydrates, lipids, urea, electrolytes
- Source: largely fetal urine in late gestation
- Analysis: amniocentesis
- Uses:
- Fetal genetic cell analysis for defects
- Chemical markers (e.g., fibronectin) to assess fetal lung maturity
Saliva
- Characteristics: clear, alkaline, viscous; produced by salivary glands
- Uses: microbial studies (viruses/bacteria), chemical tests (hormones, drugs, abused substances); blood group antigen secretor testing
- Collection: typically chewed wax or dental cotton for several minutes
Human Chorionic Gonadotropin (hCG) – Beta Subunit (Pregnancy Testing)
- What it is: pregnancy test targeting the beta subunit of hCG; also used as tumor marker in some cancers
- Specimen type: urine (first-void preferred) or serum
- Test types: qualitative (positive/negative) or quantitative (actual hCG concentration)
- Rapid strips/cassette tests: high specificity (~98\%) for hCG
- Principle: two lines indicate result; presence of test line with antibody–hCG reaction = positive
- Procedure (urine/serum):
- Remove strip from pouch; immerse in specimen without exceeding the max line
- Place on a non-absorbent surface; start timer; read at ~3\, ext{min} (urine) or 5\, ext{min} (serum)
- Interpret with trained personnel
- Quality control for urine pregnancy tests:
- External QC: test with positive and negative controls when opening a new box; document lot # and expiry; mark box as passed/ref passed
- Internal QC: presence of T line (test) and C line (control) on each strip; if C line present and T line present => valid result; if only C line present or T line absent => invalid
- Blood vs urine testing:
- Advantages of blood: earlier detection (≈3–10 days post-conception); can measure concentration for monitoring
- Disadvantages: more expensive, slower results, requires clinical lab
hCG – Beta Subunit: Procedure Details
- Pregnancy tests by strips/cassette: qualitative urine/serum detection; 98% specificity
- Reading results: based on appearance of lines after specified minutes
- QC and interpretation emphasize external and internal controls
Body Fluid Retention, Storage, and Disposal
- Retention and storage:
- Refrigerate most body fluids unless SOP specifies otherwise
- Retention times vary by specimen type (roughly 48 hours to 7 days depending on fluid and tests)
- Disposal:
- Use leak-proof, tear-resistant biohazard bags/containers; clearly labeled with biohazard symbol
- Ensure containers are sealed to prevent leakage
Quick reference values and concepts (summary)
- CSF normal adult volume: 90\text{--}150\,\text{mL}; protein ~35\,\text{mg/dL}; glucose ~60\,\text{mg/dL}; WBC count ~0--8\times 10^9/\text{L} in adults
- Fluid/serum tests differentiate transudates vs exudates using protein LDH ratios, WBC counts, and SAAG
- Synovial fluid normal WBCs: < 200\,/\text{mL}; crystals absent; high viscosity due to hyaluronate
- Sperm count reference: 20--160\times 10^6/\text{mL}
- Amniotic fluid used for fetal genetic and maturational markers
- hCG testing: qualitative or quantitative; strip tests provide rapid results; QC essential