Body fluid analysis in clinically approved

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1. Body fluid analysis in
clinical lab

2. •Collection
•Turn around time
•Physician/laboratory communication
•Reliable reference values

3. •CSF
•Synovial fluid
•Peritoneal fluid
•Pleural fluid
•Pericardial fluid

4. CSF
•10 to 60 ml……..90 to 150 ml
•Ultrafiltration and secretion
physical support
protective effect against sudden changes in acute
venous (respiratory and postural) and arterial blood
pressure
excretory waste function
maintains CNS ionic homeostasis

5. REASONS TO TEST
•Meningitis
•SAH
•Malignancy
•Demyelinating disease

6. Specimen collection

7. Required
Opening CSF pressure
Gross Examination
Total cell count and differential
Glucose (CSF/plasma ratio)
Protein
Optional
Cultures, gram stain, antigens, cytology
Protein electrophoresis, VDRL, D-dimers

8. Gross
•Crystal clear, colorless, like water
•Turbidity (WBC>200/μlor RBC>400/μl), bloody
(RBC>6000/μl)
•Viscous samples

9. Xanthochromia

10. Traumatic tap
•Decreasing amounts of
blood(Last tube will
have less)
•Clot present
SAH
•All tubes uniform
•No clot
•Hemosiderin/
hematoidinpigment

11. Microscopic examination
Total leucocyte count:
•7% neutrophils with normal WBC
•WBCcorr= WBCobs−WBCadded
•WBCadded= WBC(BLD )×RBC(CSF)/ RBC(BLD)
Adults Neonates
0-5 cells/μl 0-30 cells/μl

14. Where to count
•a) < 200 cells are present in all nine squares, count all
nine squares. This area counted is 9 mm2.
•b) > 200 cells are present in all nine squares, then
count the four corner squares. This area counted is 4
mm2.
•c) > 200 cells are present in one square, then count
five of the squares within the centersquare for an
area of 0.2 mm2

16. N X Dilution factor
Area of total squares counted X Depth
Total cell count=
Calculation of Cell count
Correlation of cell count with cytomorphological findings is
essential.

18. Slide for DLC
•Cytocentrifuge, 500g for 5 min
•2 drops of 22% albumin
•Wrights stain

23. Parasites

28. Synovial fluid
Imperfect ultra filtrate of plasma combined with
hyaluronic acid
FUNCTIONS
•Lubricate
•Provide nutrients
•Remove debris
REASONS TO TEST
•Sepsis
•Hemorrhage
•Crystal induced inflammation

29. Specimen collection

30. GROSS
•Color
•Volume
•Clarity
LOSS OF CLARITY
•Leucocytes
•Crystals
•Rice bodies
•Onchronosis

32. Lymphocytes:
•15% of SF cells
•RA and other collagen disorders, chronic infections.
Monocytes & macrophages
•Most common cells present in normal SF, 65% of the
cell count.
Eosinophilia
•>2% of the leukocyte count,
•Rheumatoid arthritis, rheumatic fever, metastatic
carcinoma, Lymesdisease, parasitic infection, chronic
urticaria, and angioedema & following arthrography.

33. Crystals
•Neutrophil predominant collection
•Intracellular crystals –pathognomic
Monosodium uratemonohydrate
Calcium pyrophosphate dihydrate
Apatite & basic calcium phosphates
Calcium oxalate
Lipids

34. Type of crystal Sizeand shape Associated conditions
Monosodium urate
(MSU)
needle-shaped rods
5–20 μm
strongly birefringent
UrateGout
Occin infections
Calcium
pyrophosphate
dihydrate(CPPD)
rhomboids, rods, or
rectangles 1–20 μm
birefringent
Hypomagnesemia,
Hemochromatosis,
Hyperparathyroidism &
Hypothyroidism
BCP small and
nonbirefringent
Non specific
Calcium oxalate
dihydrate
5-to 30-μm bipyramidal
octahedral “envelopes”
variable birefringence
chronic renal dialysis &
primary oxalosis, inborn
error of metabolism
Lipid crystals 1-to 20-μm spheres with
a Maltese cross
birefringent
Non specific
Can cause acute arthritis
Cholesterol crystals irregular birefringent
plates, notched corners
chronic effusions (e.g.,
TB arthritis, RA, SLE)

36. Crystal mimics
•Glove powder
•Talc
•Anticoagulants
•Prosthetic fragments
•Dust particles
•corticosteroids
•

37. Serous fluids
•Pleural
•Peritoneal
•Pericardial

38. Reasons to test
•Detect sepsis
•Malignancy
•Systemic disease

39. Specimen collection

40. Gross
•Transudates-usually clear
•Exudates –milky, turbid, bloody lights criteria
supernatant clear turbidity persists
cellular elements chylousor pseudochylous
centrifuge

41. Serous fluids
•Leucocytes
•RBC
•Mesothelialcells
•Macrophages(mononuclear phagocytes)
•Vacoulatedhistyocytes( can be confused with signet
ring cells)
•Bacteria
•Yeast

42. Mesothelialcells
•Distinguishable cell
borders
•Flat sheets
•Individual cells
•uniform
Malignant cells
•Poorly defined cell
borders
•Ball like clusters
•Cannibalism
•Non uniform

43. •Smooth nuclear
membrane
•No clefts
•Vacoulesare limited to
cytoplasm
•Irregular nuclear
membrane
•Clefts and moulding
•Vacoulesare all over
including over nuclei

45. DLBCL

46. CARCINOMA

47. YEAST, SPORES

48. Automation
Why automation
Labourintensive
Time consuming
Skilled person(24x7)
Interobservervariability
Biohazard

49. Problems with automation
•Carryover
•Background counting
•Flagingof abnormal cells
•Clogging of machine

50. •Cytochemistry
•Immunocytochemistry
•Flow Cytometry
•Molecular studies
Ancillary Techniques

51. Time
delay
viability
30 min 50%
60 min 20%
90 min 10%
CSF for flow cytometry
Dux et al, J Neural Sci, 1994;121:74-78

52. References
•KarcherDS and McPherson RA. Cerebrospinal, Synovial, Serous Body Fluids, and
Alternative Specimens in Henry's Clinical Diagnosis and Management by Laboratory
Methods. Eds. McPherson RA and PincusRA. Twenty Second Edition. Philadelphia, PA:
WB Saunders 2012: 480-506.
•de GraafMT, de JongsteAHC, KraanJ, BoonstraJG, SillevisSmittPAE, GratamaJW.
Flow cytometriccharacterization of cerebrospinal fluid cells. CytometryPart B 2011; 80B:
271–281.
•Sandhauset al (2010) 'Automated Cerebrospinal Fluid Cell Counts Using the SysmexXE-
5000',Am J ClinPathol,(134), pp. 734-738.
•Grimaldiand Scopacasa(2000) 'Evaluation of the Abbott CELL-DYN 4000 Hematology
Analyzer',Am J ClinPathol,(113), pp. 497-505.
•KRISHAN ET AL (2006) 'Detection of TumorCells in Body Cavity Fluids by Flow
Cytometricand ImmunocytochemicalAnalysis',Diagnostic Cytopathology,34(8), pp. 528-
541.