Re: vital stain

From:Barry Rittman

Frederick
Thank you for this excellent suggestion.
It is true that the eosin stain will fade with the correct "fluorescence"
illumination.  I would recommend that the section be examined by indirect (epi-)
illumination. The level of light intensity that is required for such
illumination is significantly lower than for transillumination. This results in
a much slower rate of fading of the eosin.
I routinely examine old H and E stained section with this and the eosin is much
more brilliant than when examined by routine brightfield microscopy..
An additional advantage is that  there is improved  resolution.
Of course not everyone has the availability of epi- illumination for their
microscope.
Barry

"Monson, Frederick C." wrote:

> Morning,
>         Just on the chance that this would be appropriate for your effort, I
> will make a suggestion for a simple, not quite-non-destructive, method for
> spotting early capillary damage in routine specimens.
>         Background:  Fresh Eosin preparations have a green sheen when
> exposed to the UV in daylight or even fluorescent illumination.  FRESH!
>
>         Utility:  As a consequence of the eosinophilia of RBC's, one can use
> any fluorescent scope with a FITC cube or filters to demonstrate the
> distribution of RBC's in any H&E section.
>
>         Downside:  After exposure to UV the eosin background will have a
> visible hole in it when viewed with normal brightfield illumination.
>
>         Upside of Downside:  One can always relocate those areas on which
> one paused 'for a look' while using the UV.
>
> Hope this helps,
>
> Fred Monson
>
> Frederick C. Monson, PhD
> Center for Advanced Scientific Imaging
> West Chester University
> West Chester, Pennsylvania, USA, 19383
> 610-738-0437
> fmonson@wcupa.edu
>
> > ----------
> > From:         Zubovits, Dr. Judit
> > Sent:         Wednesday, January 9, 2002 7:19 PM
> > To:   'histonet@pathology.swmed.edu'
> > Subject:      vital stain
> >
> > Hello Histonet-ters;
> >
> > I wonder if you could help me with the following:
> >
> > I am looking at "muscle biopsies" from volunteers (mice) who have received
> > focused ultrasound treatment.  Of course enough excess heat would cook any
> > tissue, but I we are trying to detect much more subtle changes/damage
> > produced by a lot less heat.
> >
> > I know TTC has been used on hearts to detect ischemic necrosis associated
> > with myocardial infarction, but as I understand it, you need to soak the
> > tissue *before* paraffin embedding, and TTC is intended to show areas of
> > necrosis at the gross, not microscopic level.
> >
> > Is there anything out there for the microscopic level that would highlight
> > areas of damage?
> >
> > Thanks for all your help.
> >
> > Judit Zubovits
> >
> >





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