RE: [Histonet] viability stain for 450 micron brain slices

From:"Liz Chlipala"

Try a live cell/dead cell kit. You see this application used alot on
thick sections of cartilage, etc.  I think I might have a reference for
thick sections of articular cartilage.  If you can get away without
using fluoresence I would try TTC staining.  Just search for TTC and rat
brain or something like that, its used quite a bit in the literature.
You might also try NADH.  I have a protocol for the NADH stain but its
for frozen sections and it may work for thick sections. 


Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, CO 80308
phone (303) 735-5001
fax (303) 735-3540
Ship to Address:
Premier Laboratory, LLC
University of Colorado at Boulder
MCDB, Room A3B40
Boulder, CO 80309

-----Original Message-----
[] On Behalf Of Tarango,
Sent: Wednesday, November 07, 2007 5:31 PM
Subject: RE: [Histonet] viability stain for 450 micron brain slices

The first thing that comes to mind is Trypan blue, but that would be for
regular light microscopy.  If you want to do florescent you might try
7AAD or propidium idodide.  An ethidium bromide / acridine orange combo
might be good too.  

I'm not sure about fixing the staining.  

Mark Adam Tarango HT(ASCP)
Histology & IHC Supervisor
Nevada Cancer Institute
One Breakthrough Way
Las Vegas, NV  89135
Direct Line (702) 822-5112
Treo (702) 759-9229
Fax (702) 939-7663


-----Original Message-----
[] On Behalf Of
Sent: Wednesday, November 07, 2007 6:44 AM
Subject: [Histonet] viability stain for 450 micron brain slices

I am looking for a staining technique to test the viability of slices of
the rat brain after some force-displacement tests on the tissue. As I am
an engineer and no chemist I do not know what kind of stain would be
appropriate. We keep the slices from the cerebral cortex and the corpus
callosumalive alive in a tissue chamber with ACSF that is bubbled with
95% O2/ 5% CO2. The slices have a thickness of 450 micron and there is a
fluorescent confocal microscope available. As this microscope is away
from the testing site and we do not have the equipment that can keep the
tissue alive during imaging, it would be very convenient to fix the
tissue after staining. Does anyone of you have experience with a
staining technique that is suitable for my experiment?

Best, Kirsten Henken

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