Tag Archives: Electron Microscopy

Preparing cells for Flow Cytometry and Electron Microscopy

Cells from experiments that are to be analyzed via Flow Cytometry or Electron Microscopy are prepared in the following manner:

Flow Cytometry:

Harvest cells and centrifuge (1000 rpm, 3 min),  vortex the cell pellet  with .5mL of cold PBS. Add another .5mL of cold PBS  and completely resuspend the pellet.  Centrifuge the cells again.  Vortex the cell pellet was  with .2mL of cold PBS and add the cells dropwise to .750mL of cold 75% ethanol. Incubate the cells on ice for 30 minutes before being transferring to the fridge. Cells can be stored in this fashion for up to 1 week.

Electron Microscopy:

Harvest 1.5 mL of cells and centrifuge off media (1000 rpm, 5 min). Wash twice with 500 uL of PBS. Fix cells with 150 uL of either 2% paraformaldehyde or cold 75% ethanol for 10 minutes in the fridge. Resuspend cell pellet in PBS.