Quantitative analysis of leukocyte oxidative burst in 100 µl of whole blood under optimal conditions without any disturbance by isolation procedures A. Sample Preparation for low and high stimulant. Simple as you know it from immunofluorescence analysis: 1. Activation 2. Oxidation 3. Lysing and Fixation 4. Washing and DNA Staining B. Measurement Gating on leukocytes after DNA staining: Discrimination of bacteria, single leukocytes and doublets C.Data Evaluation Gate on granulocytes and determine the percentage of oxidizing cells and their enzymatic activity (= mean fluorescence = number of cleaved substrate) Report your results

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