Synchronised phagocytosis is used to ensure that all observed phagocytosis events in a maple have occurred after a similar amount of time. This is useful for microscopy and molecular biology, when discrete stages of phagocytosis want to be investigated.
- Prepare phagocytes and target particles as per usual.
- Remove phagocytes from incubator and allow to cool to room temperature.
- Add the desired number of target particles, and centrifuge sample for 1 min at 1500 RPM (~500xG).
- Let sit at room temperature for 15-20 min. During this time phagocytes will bind particles, but not ingest them.
- Wash phagocytes 2-3X with PBS or media to remove unbound target particles.
- Activate phagocytosis by swapping media for media warmed to 37oC, then transfer to an incubator for the desired amount of time.
- Terminate phagocytosis by transferring cells to ice, adding a fixative, or lysing cells.