[Histonet] RE: Section cells on a plastic trans well membrane

[Elizabeth Chlipala]

Jamie

We have sectioned these before and we used charged slides and did not have a problem with section adherence.  I would fix longer than 5 minutes.  We would use biopsy punches to remove the membrane from the well we would process the disc whole between biopsy pads and bisect prior to embedding.  You need to use a good plus slide and it you are still having problems I would let the unstained slides sit for a couple days prior to staining.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
PO Box 18592
Boulder, CO 80308
(303) 682-3949 office
(303) 881-0763 cell
(303) 682-9060 fax
liz <@t> premierlab.com

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Premier Laboratory, LLC
1567 Skyway Drive, Unit E
Longmont, CO 80504
________________________________________
From: histonet-bounces <@t> lists.utsouthwestern.edu [histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Erickson, Jamie E [jamie.erickson <@t> abbvie.com]
Sent: Tuesday, July 09, 2013 2:52 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Section cells on a plastic trans well membrane

Hi All,
                  Has anyone tried to section cells cut from a trans well dish? We have cells that are cultured and attach to a trans well membrane and our pathologist wants to look at them on the membrane but the membrane is a thin plastic film...see the problem.
We fixed the cells in the trans well for 5 minutes then  cut the trans well out of the well and bisected it and processed overnight we use paraffin  type #1 paraffin. The next morning we embedded it with type #9 paraffin.  The samples cut but  do not stay on the slide after xylene. I think the plastic does not adhere to the slide. Would a adhesive slide work to keep the thin plastic stuck to the slide??

My pathologist does not want to change to a collagen membrane  so he wants me to find another (harder paraffin) to use, I don't think that will help.
Has anyone does similar processing/embedding?
I'll take any help I can get..

Thanks,

Jamie
Abbvie Bioreseach Center
Pharmacology
100 Research Dr.
Worcester,Ma 01605
OFFICE    +1 508-688-3134
FAX          +1 508-793-4895
EMAIL  jamie.erickson <@t> abbvie.com<mailto:jamie.erickson <@t> abbvie.com>


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