New discovery suggests a novel strategy against harmful inflammation NatureComms
. In this method, the position of individual molecules is localized with high accuracy by switching them on and off sequentially using appropriate laser power settings and buffer conditions. For neutrophil experiments, cells seeded in glass bottom dishes are fixed with freshly made 4% paraformaldehyde for 10 min at room temperature, permeabilized, and blocked for 1 h with 1% BSA in PBS.
using default settings, resulting in a single image with improved resolution , in which individual granules could be visualized. At least 5 fields-of-view were acquired for each condition in 3 separate experiments.Mouse neutrophils isolated from the bone marrow of WT andmice, were resuspended in phenol-red free RPMI and either treated with GM‐CSF , LPS , cytochalasin D or left untreated for 30 min at 37 °C, at which point the cells were stimulated with 10 µM fMLF for 10 min.
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