Background: We describe a method for counting leukocytes in the Nageotte hemocytometer by fluorescence microscopy. Material and Methods: Leukocyte nuclei were stained with a DNA fluorochrome. Hemoglobin interferes absorbing the light emitted by the stained nuclei. In order to avoid this, the sample was diluted with a stromatolysing reagent and then centrifuged on a density gradient. Leukocytes were recovered in the pellet, while most of hemoglobin and red blood cell stromata were discarded with the supernatant. At the end, the sample was concentrated 2 times. Results: The absolute limit of sensitivity was 0.01 leukocytes/ml. Counting efficiency was measured and found to be 87.1% on average (range 35.7–135.5%). Conclusions: Counting is much easier and faster than with usual methods and less prone to subjective biases. The method is also valid for components stored more than 4 weeks.

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1999
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