Clinical Diagnostic for the Quantitative Analysis of Leukocyte Phagocytosis in Human Whole Blood
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A. Sample Preparation
or opsonized E.coli-FITC.
Simple as you know it from immunofluorescence analysis:
1. Labeling  | 2. Quenching  |
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3. Lysing and Fixation  | 4. Washing and DNA Staining  |
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B. Measurement
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Gating on leukocytes after DNA staining:
Discrimination of bacteria, single leukocytes and doublets
C. Data Evaluation
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Gate on monocytes and granulocytes and determine the percentage of phagocytosing cells and their phagocytic activity (= mean fluorescence = number of bacteria per cell).