FM4-64 staining protocol                                                                                                                                           060427 Toshiyuki Hatano

Grow cells to log phase in 30C°
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Add FM-64 (final 1-1.5 μM) in the culture medium
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Incubate in 30 C° for 15 mini
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Concentrate cells by centrifugation 3000 rpm 5 min and re-suspending the pellet with small amount of medium (~100μl)
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Put the cell on 0.1% poly-L-lysine coated slide glass
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Stand for 10 minutes
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Wash by pipette suspension for 5 times with 100μl M9 medium FM-64 (final 1-1.5 μM)
 x2
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mount with the M9 medium +FM-64 (final 1-1.5 μM)