Re: Vital marker
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From: | Paul Millikin <millikin@mtco.com> (by way of Histonet) (by way of histonet) |
To: | histonet <histonet@magicnet.net> |
Reply-To: | |
Content-Type: | text/plain; charset="us-ascii" |
Ian (by way of Histonet) wrote:
>
> Is anyone aware of an intracellular marker that is ingested (or absorbed) or
> somehow gets into cells in culture media? This marker needs to be detectable
> with light microscopy. I know of fluorescein labels but I can't use
> fluorescence microscopy because of the high collgen content of the substrate
> the cells are being seeded onto. Any ideas would be appreciated. Thanks, Ian
> Birchall.
Have not tested it in cell culture, but toluidine blue makes a great
supravital stain for blood and other body fluids. See Millikin, PD, "A
Supravital Stain for Nucleoli in Human Lymphocytes," American Journal of
Clinical Pathology, 1974:62;520-24.
0.25-0.50% toluidine blue in 100% ethanol. Warm clean slides at 60%C,
dip in the alocholic solution, and stand on end to drain and dry. Slides
are good for months. Put on a SMALL drop of fluid, add coverslip, and
wait 2-5 minutes to allow for stain uptake. Preparation is not permanent
because the cells are destroyed after 20-30 minutes. They are
photogenic, though, if a permanent record is desired.
For whatever it's worth.
--
Paul Millikin
Peoria, IL
millikin@mtco.com
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