RE: [Histonet] in situ question

From:Rene J Buesa



Exactly so! When you add water to the solid formaldehyde polymerá(=paraformaldehyde) you are just preparing a formaldehyde solution.
RenÚ J.

--- On Tue, 8/19/08, Tony Henwood  wrote:

From: Tony Henwood 
Subject: RE: [Histonet] in situ question
To: "N Fournier" , histonet@lists.utsouthwestern.edu
Date: Tuesday, August 19, 2008, 8:01 PM

Some questions,

What is "PARA"?

Is it an something you add to the formaldehyde solution?

Please don't use the term 4% paraformaldehyde, use 4% formaldehyde,
unless you are rolling the tissue in a powder consisting of 4%
paraformaldehyde and 96% talcum powder or some other solid!

Please refer to: Manoonkitiwongsa & Schultz (2002) Histochem J 34:
365-367



Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145 




-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of N
Fournier
Sent: Wednesday, 20 August 2008 3:37 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] in situ question


Hi,

I have a few questions regarding in situ hybridization. The protocol
that I am following indicates that they perfused rats with PBS followed
by 4% paraformaledhyde (in PBS). However, I always thought that one
generally extracts the rat brain fresh, flash freeze, section, and then
post-fix in 4% paraformaldehyde before commencing with the in situ
hybridization. I suppose it might not matter since in the end the tissue
is post-fixed in PARA; however, I would like to hear what others with
experience doing in situ think about this procedure. Additionally, does
one always use DEPC treated solutions even for these perfusion steps
(e.g., PBS perfusion, paraformaldehyde perfusion)? Lastly, does one have
to section on a cryostat? Could I simply use a vibrating microtome to
section the tissue instead. 

Thanks for the help everyone,

Neil


_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet

*********************************************************************
This email and any files transmitted with it are confidential and intended
solely for the use of the individual or entity to whom they are addressed. If
you are not the intended recipient, please delete it and notify the sender.

Views expressed in this message and any attachments are those of the individual
sender, and are not necessarily the views of The Children's Hospital at
Westmead

This note also confirms that this email message has been
virus scanned and although no computer viruses were detected, The Childrens
Hospital at Westmead accepts no liability for any consequential damage resulting
from email containing computer viruses.
**********************************************************************


_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet
      
_______________________________________________
Histonet mailing list
Histonet@lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet




<< Previous Message | Next Message >>