Downloaded 85 times
Uploaded byRavi Kumudesh
Haemocytometry by Ravi Kumudesh
The document provides a comprehensive overview of hemocytometry, focusing on the measurement of red and white blood cells (RBCs and WBCs) using various techniques and equipment such as hemocytometers and pipets. It includes reference ranges for RBC and WBC counts, examples of dilutions, and counting rules for accurate results. Additionally, it emphasizes the proper care and handling of microscopes used in these measurements.
More Related Content
Similar to Haemocytometry by Ravi Kumudesh
Haemocytometry by Ravi Kumudesh
- 1. Haemocytometry Ravi Kumudesh MSc/BSc/Dip(MLT) Sri LankaSociety for Medical Laboratory Science 1
- 2. Hemocytometry • Hemocytometer • RBCand WBC • Thoma pipet • Unopette • Dilution problem • Microscopy 2 Sri Lanka Society for Medical Laboratory Science
- 3. Red Cell andWhite Cell Counts • RBC problem: anemia, polycythemia • WBC problem: leukocytosis, leukopenia • RBC reference ranges • Male 4.50 - 6.00 x 106 /µL (x 1012 /L) • Female 3.70 - 5.20 x 106 /µL (x 1012 /L) • WBC reference range • Both 5.0 - 10.0 x 103 /µL (x 109 /L) 3 Sri Lanka Society for Medical Laboratory Science
- 4. Diluting Fluid • ForRBC – Physiologic saline (0.8% NaCl) – Hayem’s solution – Reese-Ecker • For WBC – 3% Acetic acid – 0.1 N HCl 4 Sri Lanka Society for Medical Laboratory Science
- 5. Thoma Pipet • RBC –Aspirate to 0.5 or 1.0 – Dilute to 101 – DF = 200 or 100 • WBC – Aspirate to 0.5 or 1.0 – Dilute to 11 – DF = 20 or 10 • Discard volume of one (1) in the stem 5 Sri Lanka Society for Medical Laboratory Science
- 6. 6 Sri Lanka Societyfor Medical Laboratory Science
- 7. Improved Neubauer Hemocytometer 7 SriLanka Society for Medical Laboratory Science
- 8. Hemocytometer Chamber • Depth= 0.1 mm • Grids – RBC use 5 small squares in the center large square – WBC use 4 corner large squares – Unopette for WBC use all 9 large squares 8 Sri Lanka Society for Medical Laboratory Science
- 9. Hemocytometer Chamber 9 Sri Lanka Societyfor Medical Laboratory Science
- 10. Counting Rule • Donot count cells touching – Bottom line – Right line 10 Sri Lanka Society for Medical Laboratory Science
- 11. 11 Sri Lanka Societyfor Medical Laboratory Science
- 12. 12 Sri Lanka Societyfor Medical Laboratory Science
- 13. RBC Example… • Aspirateblood to 0.5, then dilute to 101 • DF = 200 • Counted 500 erythrocytes in 5 small squares RBC = 500 rbc x 25 Sm Sq x DF (200) 5 Sm Sq 0.1 mm3 = 5.00 x 106 /mm3 (µL) 13 Sri Lanka Society for Medical Laboratory Science
- 14. 14 Sri Lanka Societyfor Medical Laboratory Science
- 15. WBC Example… • Aspirateblood to 0.5, then dilute to 11 • DF = 20 • Counted 160 leukocytes in 4 large squares WBC = 160 wbc x 1 Lg Sq x DF (20) 4 Lg Sq 0.1 mm3 = 8.0 x 103 /mm3 (µL) 15 Sri Lanka Society for Medical Laboratory Science
- 16. Unopette • Self-contained reservoirand 20 µL pipet • Ammonium oxalate diluent • DF = 100 • Count leukocytes in all 9 large squares • Special formula to facilitate calculation 16 Sri Lanka Society for Medical Laboratory Science
- 17. Unopette 17 Sri Lanka Societyfor Medical Laboratory Science
- 18. Unopette WBC Example… •Counted 70 leukocytes in 9 large squares • Add 10% of count • Multiply by DF WBC = (70 wbc + 7 wbc) x DF (100) mm3 = 7.7 x 103 /mm3 = 7.7 x 109 /L 18 Sri Lanka Society for Medical Laboratory Science
- 19. Compound Microscope • Ocular (adjustable binocular) •Mechanical stage • Condenser • Objectives (parfocal) 19 Sri Lanka Society for Medical Laboratory Science
- 20. Care of Microscopes •Carry with care • Protect long objective lens • Clean and disinfect stage • Wipe oil off OIL • Save lamp life 20 Sri Lanka Society for Medical Laboratory Science
- 21. Sri Lanka Societyfor Medical Laboratory Science 21