1 / 19

Week 4: Hemocytometry

Hemocytometer RBC and WBC Thoma pipet Unopette. Dilution problem Microscopy. Week 4: Hemocytometry. Red Cell and White Cell Counts. RBC problem: anemia, polycythemia WBC problem: leukocytosis, leukopenia RBC reference ranges Male 4.50 - 6.00 x 10 6 /  L (x 10 12 /L)

Rita
Download Presentation

Week 4: Hemocytometry

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Hemocytometer RBC and WBC Thoma pipet Unopette Dilution problem Microscopy Week 4: Hemocytometry

  2. Red Cell and White Cell Counts • RBC problem: anemia, polycythemia • WBC problem: leukocytosis, leukopenia • RBC reference ranges • Male 4.50 - 6.00 x 106/L (x 1012/L) • Female 3.70 - 5.20 x 106/L (x 1012/L) • WBC reference range • Both 5.0 - 10.0 x 103/L (x 109/L)

  3. Diluting Fluid • For RBC • Physiologic saline (0.8% NaCl) • Hayem’s solution • Reese-Ecker • For WBC • 3% Acetic acid • 0.1 N HCl

  4. 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 Thoma Pipet

  5. Improved Neubauer Hemocytometer

  6. 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

  7. Hemocytometer Chamber

  8. Counting Rule • Do not count cells touching • Bottom line • Right line

  9. RBC Example… • Aspirate blood 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 (L)

  10. WBC Example… • Aspirate blood 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 (L)

  11. Unopette • Self-contained reservoir and 20 mL pipet • Ammonium oxalate diluent • DF = 100 • Count leukocytes in all 9 large squares • Special formula to facilitate calculation

  12. Unopette

  13. 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

  14. Compound Microscope • Ocular (adjustable binocular) • Mechanical stage • Condenser • Objectives (parfocal)

  15. Care of Microscopes • Carry with care • Protect long objective lens • Clean and disinfect stage • Wipe oil off OIL • Save lamp life

More Related