Synchronised Phagocytosis

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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.

  1. Prepare phagocytes and target particles as per usual.
  2. Remove phagocytes from incubator and allow to cool to room temperature.
  3. Add the desired number of target particles, and centrifuge sample for 1 min at 1500 RPM (~500xG).
  4. Let sit at room temperature for 15-20 min.  During this time phagocytes will bind particles, but not ingest them.
  5. Wash phagocytes 2-3X with PBS or media to remove unbound target particles.
  6. Activate phagocytosis by swapping media for media warmed to 37oC, then transfer to an incubator for the desired amount of time.
  7. Terminate phagocytosis by transferring cells to ice, adding a fixative, or lysing cells.