Changes

FM4-64 Labeling of Yeast Cells

453 bytes added, 16:47, 7 December 2009
added details from Emily
==Materials==
*Instead of FM4-64 we use SynaptoRed (Sigma #S6689)
*Media (typically [[YPDA Media]])
*Dye solution. For each cell line need 250 uL media + 3 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.#Quench Spin down cells for and wash 2x with media.#Add cells to 5 mL media and mix. Take a 1 mL sample and measure A600#Incubate 3hwith 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]]