[Histonet] Polyester membrane

[Kim Burns]

Hi Patsy,

On which type of membrane are the cells grown? A transwell collagen
membrane, t-clear, milli cell, snap well?

We process cells grown on these types of polyester membranes on a regular
basis. We leave the membrane attached to the support and process as usual,
the exception being that we do not use xylene on our processor. We use a
substitute called slide brite. The xylene can be to harsh for the support.
After processing we sandwich the membrane between layers of paraffin and
then cut the membrane from the support. We then cut the membrane in half,
stand it on end in a mold and make the paraffin block. 

The best membranes to work with are transwells. T-clears tend to separate
from the paraffin making sectioning difficult and have a tendency to come
off subbed slides taking the cells with them.

Kimberlie Burns
Cystic Fibrosis Research Center
University of North Carolina









Date: Tue, 12 Oct 2004 11:03:13 -0600
From: "Patsy Ruegg" <pruegg <@t> ihctech.net>
Subject: [Histonet] Polyester membrane
To: "'James Watson'" <jwatson <@t> gnf.org>,	"'Kim Merriam'"
	<kmerriam2003 <@t> yahoo.com>,	"'Histonet'"
	<histonet <@t> lists.utsouthwestern.edu>
Message-ID: <001801c4b07d$5bf7e5f0$83020a0a <@t> IHCTech>
Content-Type: text/plain;	charset="us-ascii"

Has anyone ever used HISTOGEL as a freezing embedding media?  Also, I was
wondering what experiences if any you all have had with processing for
sectioning polyester membranes with cells grown on them.  Frozen, Paraffin,
GMA?????  I am securing the cut out membrane in histogel on edge and
thinking of trying to gently paraffin process and/or freeze section??? Patsy