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What happens in the aperture?

Learn about hematological cell counting, differential cell discriminators, impedance cell counting principles, and hemoglobin measuring techniques under a microscope. Explore the essential parameters and methods involved in analyzing blood samples.

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What happens in the aperture?

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  1. What happens in the aperture? Basics of Hematology cell counting

  2. Blood cells

  3. NRBC EO RBC RBC NEU MONO LYM BASO PLT Blood cells – under microscope

  4. Hematology parameters 1

  5. Hematology parameters 2

  6. 1.: RBC-LYM discriminator 2.: LYM-MID discriminator 3.: MID-GRA discriminator 1. 2. 3. Hematology parameters 3

  7. Blood sample

  8. Impedance cell counting principle dR/R = 1 / 20 000 = 50ppm, RBC = 1mV, min PLT= 40µV

  9. Aperture clogging effect

  10. Coincidence effect • >1 cells in aperture look one big cell • Less counts, distorted histogram • Solution: diluting samples, mathematical coincidence correction Linearity range (80um): WBC: 100 x 10^3/ul RBC: 10 x 10^6/ul PLT: 1000 x 10^3/ul

  11. 3. Hemolyser 1. 2. Blood 1:32.000 1:200 Sample preparation: diluting, lysing • a + diluent → b • b + diluent → c → RBC/PLT • b + Lyser → WBC/HGB a b c

  12. PLT & RBC LYM MON GRAN Differential Lysing process RBCs destroyed WBCs selectively shrank to nuclei

  13. & HGB 3-part Differential WBC Histogram

  14. Hemoglobin measuring principle • Specific wavelength light source – green LED @ 560 nm • Light to frequency converter • Ambient light compensation with U/D counting, and light chopping LED SAMPLE DETECTOR LED switching: Frequency output

  15. Hemoglobin calculation – HGB blank • HGB needs blank measurement on clean diluent to compensate temperature drift HGB = C * ln (CNTblank / CNTsample) CNTblank = HGB count on diluent = 10.000 CNTsample = HGB count on sample = 4.000

  16. Thank you for your attention!

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