The fluorescence was compared with controls in 5,000 events. The phagocytosis rate (%) was calculated according to the following formula: phagocytosis (%) = 100 × [fluorescent cells (sample—negative control) / fluorescent cells (positive control – negative control)]. Positive and negative controls were the addition of fluorescent beads and DMSO alone, respectively.
Quantifying Phagocytosis in Macrophages
The fluorescence was compared with controls in 5,000 events. The phagocytosis rate (%) was calculated according to the following formula: phagocytosis (%) = 100 × [fluorescent cells (sample—negative control) / fluorescent cells (positive control – negative control)]. Positive and negative controls were the addition of fluorescent beads and DMSO alone, respectively.
Corresponding Organization : Yasuda Women's University
Other organizations : University of Basel, International University of Health and Welfare
Variable analysis
- Concentration of kinase inhibitors (128 to 0.03 μM)
- Phagocytosis rate (%)
- Cell type (J774.1 or THP-1 macrophages)
- Incubation time (2 h)
- Bead size (2 μm diameter)
- Opsonization of beads (10% human serum)
- Multiplicity of infection (MOI of 10)
- Positive control: Addition of fluorescent beads
- Negative control: Addition of DMSO alone
- Cytochalasin D (10 μM) as a control to completely inhibit actin polymerization
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