Quantitative determination of the chemotactic function of neutrophilic granulocytes
A. Sample Preparation
1. Leukocyte Isolation
2. Stimulation and Chemotaxis
3. Cell labelling
4. DNA Staining, Measurement
B. Measurement
Acquire data by using fluorescence triggering in the FL3 channel. Region 1 set on leukocytes and counting beads
C. Data Evaluation
Region 2 is set on counting beads during data acquisition. Set a logical gate on granulocytes in region 1 and region 3 and analyse the number of cells and the Mean value of the FSC signal for the population of granulocytes.
Determine the percentage of activated neutrophiles showing a decreased expression of the cell adhesion molecule LECAM-1 (separate FL1 histogram).
