Re: Vital marker

<< Previous Message | Next Message >>
From:Paul Millikin <> (by way of Histonet) (by way of histonet)
To:histonet <>
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

<< Previous Message | Next Message >>