Reply: Fluorescent dyes

Sat Jul 24 19:37:08 EST 1993

DiOC6 has shorter hydrocarbon "tails" than DiI. This feature makes it  
more soluble but also more "promiscuous".  It is very easy to label  
living cells - just add a microliter of 10mg/ml DiOC6 in ethanol to a  
milliliter of cell suspension (at 10super7 cells per ml).  As quickly  
as you can get a slide ready, the cells are labeled.  However, the  
DiOC6 doesn't stay in the membranes as DiI does.  If we wash cells  
out of excess DiOC6, and add unlabeled cells, all the cells become  
labeled.  We think that what we see represents an equilibrium of dye  
moving in and out of various lipophilic compartments in the cell. The  
nice thing about this equilibration is that the labeling is quite  
stable.  When a dye molecule is bleached, another from the external  
pool can exchange with it.  

Let us know how the staining goes.

Robin Wright

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