From owner-cellbiol@net.bio.net Mon Feb 24 22:00:00 1997 Path: biosci!rutgers.rutgers.edu!uwm.edu!newsfeeds.sol.net!newspump.sol.net!howland.erols.net!rill.news.pipex.net!pipex!warwick!leicester!usenet From: "Dr E. Buxbaum" Newsgroups: bionet.cellbiol Subject: Re: vital dyes in the visible range Date: 25 Feb 1997 09:42:44 GMT Organization: University of Leicester (PCFS User) Lines: 23 Message-ID: <5euc6k$6g7@falcon.le.ac.uk> References: <330D8113.60A1@gold.tc.umn.edu> NNTP-Posting-Host: pc47.msb.le.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.22 (Windows; I; 16bit) bhadr001@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! .