View source for FM4-64 Labeling of Yeast Cells

==Materials==
*Instead of FM4-64 we use SynaptoRed (Sigma #S6689).  This is resuspended 1mg to 500 uL (2ug/mL) with DMSO and stored in '''Molecular Biology Stuff'''
*Media (see [[YPD Media and Agar]])
*Dye solution.  For each cell line need 190 uL media + 60 uL dye.

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

[[Category:Yeast]]
[[Category:Immunoflouresence]]

see http://www.mcdb.lsa.umich.edu/labs/klionsky/FM4-64labeling.pdf