FM4-64 Labeling of Yeast Cells

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  • Instead of FM4-64 we use SynaptoRed (Sigma #S6689). This is resuspended 5mg to 514 uL (16mM) with DMSO and stored in Molecular Biology Stuff
  • Media (see YPD Media and Agar)
  • Dye solution. For each cell line need 200 uL media + 1 uL dye.


  1. Grow and overnight culture of yeast in 4 mL of the appropriate media.
  2. Spin cells down and resuspend in dye solution FM4-64. Wrap in alluminum foil and incubate with shaking at 24C for 1h.
  3. Spin down cells and wash 2x with media.
  4. Add cells to 5 mL media and mix. Take a 1 mL sample and measure A600
  5. Incubate 3h with shaking at 24C. Take a 1 mL sample and measure A600. This should be one full doubling from the previous reading.
  6. Mount cells on coverslip and analyze by fluorescent microscopy