digest reply (Fekete's/MIB-1/Cryosections

<< Previous Message | Next Message >>
From:Melody Ricci <melodyricci@usa.net>
To:Histonet Mailing List <histonet@Pathology.swmed.edu>
Date:Tue, 13 Jul 1999 17:18:28 -0500 (CDT)
Content-Type:text/plain; charset=US-ASCII

Hi all:

In reply to the daily digest:

Mike (Fekete's fixative):  I did a large part of my grad work using this
protocol.  The reference is very old, and I'm sure they counted them all
individually.  I utilized an image analysis system and semi-quantified based
on the pixels.  This wasn't easy since the optimum visualization occurs when
the lungs are submerged in water for viewing.  Contact me at the following
address for more specific info on this subject. I could fax you my protocols
or data if you like.  mricci@pharmingen.com or phone at (619) 812-8800 ext
3346 (San Diego, USA)

Rick (MIB-1) protocol:  Citrate Buffer pretreatment in the micro for 10
minutes followed by 20 minutes cooling on the bench.  Primary antibody
incubation overnight on paraffin sections at 4 degrees celcius.  Tonsil
positive control.

Cryosections/Breast tissue:  We often receive tissue in this way.  Although
its not optimal, we cut a chunk frozen and immediately re-freeze it in OCT
(using isopentane) to make a block for sectioning.  There is often some
freezing artifact around the edge of the tissue from it rewarming, but the
inner tissue is usually salvageable.

Melody Ricci, IHC-QC
PharMingen, Becton Dickinson Biosciences
San Diego

Get free e-mail and a permanent address at http://www.amexmail.com/?A=1

<< Previous Message | Next Message >>