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vital dyes in the visible range

Dr E. Buxbaum EB15 at le.ac.uk
Tue Feb 25 04:42:44 EST 1997


bhadr001 at GOLD.TC.UMN.EDU (kiran bhadriraju) wrote:
>Hi!
>
>I am in search of a dye to stain living cells which would absorb in 
>the in the visible range.  I looking at hepatocytes through an 
>inverted microscope (attached to an atomic force microscope, just to 
>give you the whole picture) and they don't show very well at all. 

Why not use phase contrast instead (or the poor mans substitute: oblique 
illumination)? That way you do not have to worry about ill effects of the 
dyes. Most vital stains, by the way, are fluorescent dyes. Check the 
Molecular Probes catalogue for something suitable.

How to do oblique illumination: Adjust your condensor apperture properly, 
then measure its diameter. Prepare a disk (some plastic) so that it fits 
into the filter holder of the condensor. Drill a hole into it with the 
diameter of the condensor apperture, but in such a way, that the rim of 
that hole goes through the center of the disk (with large apperture 
objectives the hole may not be completely on the disk). You will need a 
separate disk for every objective you want to use. Place the correct disk 
into the filter holder of the condensor and have a look: Quite 
spectacular!




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