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Body fluid analysis in clinically approved
- 1. Body fluid analysisin clinical lab
- 2. • Collection • Turnaround time • Physician/laboratory communication • Reliable reference values
- 3. • CSF • Synovialfluid • Peritoneal fluid • Pleural fluid • Pericardial fluid
- 4. CSF • 10 to60 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
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- 7. Required Opening CSF pressure GrossExamination 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/μl or RBC>400/μl), bloody (RBC>6000/μl) • Viscous samples
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- 10. Traumatic tap • Decreasingamounts of blood(Last tube will have less) • Clot present SAH • All tubes uniform • No clot • Hemosiderin/ hematoidin pigment
- 11. Microscopic examination Total leucocytecount: • 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 center square for an area of 0.2 mm2
- 16. N X Dilutionfactor 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
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- 28. Synovial fluid Imperfect ultrafiltrate of plasma combined with hyaluronic acid FUNCTIONS • Lubricate • Provide nutrients • Remove debris REASONS TO TEST • Sepsis • Hemorrhage • Crystal induced inflammation
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- 30. GROSS • Color • Volume •Clarity LOSS OF CLARITY • Leucocytes • Crystals • Rice bodies • Onchronosis
- 32. Lymphocytes: • 15% ofSF 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, Lymes disease, parasitic infection, chronic urticaria, and angioedema & following arthrography.
- 33. Crystals • Neutrophil predominantcollection • Intracellular crystals – pathognomic Monosodium urate monohydrate Calcium pyrophosphate dihydrate Apatite & basic calcium phosphates Calcium oxalate Lipids
- 34. Type of crystalSize and shape Associated conditions Monosodium urate (MSU) needle-shaped rods 5–20 μm strongly birefringent Urate Gout Occ in 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 • Glovepowder • Talc • Anticoagulants • Prosthetic fragments • Dust particles • corticosteroids •
- 37. Serous fluids • Pleural •Peritoneal • Pericardial
- 38. Reasons to test •Detect sepsis • Malignancy • Systemic disease
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- 40. Gross • Transudates- usuallyclear • Exudates – milky, turbid, bloody lights criteria supernatant clear turbidity persists cellular elements chylous or pseudochylous centrifuge
- 41. Serous fluids • Leucocytes •RBC • Mesothelial cells • Macrophages(mononuclear phagocytes) • Vacoulated histyocytes( can be confused with signet ring cells) • Bacteria • Yeast
- 42. Mesothelial cells • Distinguishablecell borders • Flat sheets • Individual cells • uniform Malignant cells • Poorly defined cell borders • Ball like clusters • Cannibalism • Non uniform
- 43. • Smooth nuclear membrane •No clefts • Vacoules are limited to cytoplasm • Irregular nuclear membrane • Clefts and moulding • Vacoules are all over including over nuclei
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- 48. Automation Why automation Labourintensive Time consuming Skilled person(24x7) Interobserver variability Biohazard
- 49. Problems with automation •Carryover • Background counting • Flaging of abnormal cells • Clogging of machine
- 50. • Cytochemistry • Immunocytochemistry •Flow Cytometry • Molecular studies Ancillary Techniques
- 51. Time delay viability 30 min 50% 60min 20% 90 min 10% CSF for flow cytometry Dux et al, J Neural Sci, 1994;121:74-78
- 52. References • Karcher DSand McPherson RA. Cerebrospinal, Synovial, Serous Body Fluids, and Alternative Specimens in Henry's Clinical Diagnosis and Management by Laboratory Methods. Eds. McPherson RA and Pincus RA. Twenty Second Edition. Philadelphia, PA: WB Saunders 2012: 480-506. • de Graaf MT, de Jongste AHC, Kraan J, Boonstra JG, Sillevis Smitt PAE, Gratama JW. Flow cytometric characterization of cerebrospinal fluid cells. Cytometry Part B 2011; 80B: 271–281. • Sandhaus et al (2010) 'Automated Cerebrospinal Fluid Cell Counts Using the Sysmex XE- 5000', Am J Clin Pathol, (134), pp. 734-738. • Grimaldi and Scopacasa (2000) 'Evaluation of the Abbott CELL-DYN 4000 Hematology Analyzer', Am J Clin Pathol, (113), pp. 497-505. • KRISHAN ET AL (2006) 'Detection of Tumor Cells in Body Cavity Fluids by Flow Cytometric and Immunocytochemical Analysis', Diagnostic Cytopathology, 34(8), pp. 528- 541.