[Histonet] (no subject)

[Elizabeth Chlipala]

Sylvie

We frequently embed our tissue in agarose for vibratome sections.  We
cut 30-50 micron sections of fixed mouse spleen and liver.  After the
sectioning you can carefully remove the agarose that is around the
tissue.  We then stain the 50 micron sections free floating for IF and
evaluate them with the confocal microscope. 

My question to Naira is why are you trying to cut 4 - 10 micron frozen
sections on a vibratome, why are you not cutting the frozen sections in
a cryostat.  The thinnest we can cut on the vibratome we have here is 30
microns. Sectioning in a cryostat would be your best bet.

Liz  

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
liz <@t> premierlab.com
www.premierlab.com
 
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of
Margaryan, Naira
Sent: Friday, March 25, 2005 8:45 AM
To: s.liabeuf <@t> wanadoo.fr; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] (no subject)

Other question for vibratome:

I have to do vibratome sections on frozen tissue- mouse breast with
skin. Tissue defrosting during the vibritiming and looks like fresh
tissue. It is very hard to cut. Can you give me any recommendation about
frequency, amplitude and speed? I am going to make 4-10 µ think
sections.

 

Any suggestion will be appreciate,

Naira 

 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Sylvie
LIABEUF
Sent: Friday, March 25, 2005 9:28 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] (no subject)

 

Hello,

Do anyone know if agarose used for vibratome section is a problem for
immunostaining?

thank you very much for your help

sylvie

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