[Histonet] histogel

[S Ladd]

Since your cells were fixed in formalin, perhaps there was enough residual
formalin to sufficiently fix the histogel? You use only a few drops of
histogel. My cells were not in formalin and I use about 0.5 mL of histogel.
Just for fun, you could try processing a little button of histogel (without
formalin fixed cells) and see if it dissolves?
I use ethanol and xylene on my processor and I also start with 70% ethanol.
Sharron

-----Original Message-----
From: Kyle-Byrne, Carrie - Labvision [mailto:CKByrne <@t> labvision.com]
Sent: Tuesday, April 13, 2004 11:23 AM
To: 'S Ladd'
Subject: RE: [Histonet] histogel


Sharron,
Interesting.  I've never heard of this before.  I've used Histogel for years
and never had to 'fix' it in formalin.  I used it to make cell pellets for
use as IHC control material.  I would fix the cells in formalin first then
transfer to 70% EtOH and hold them until I had several to pellet.  I'd
decant the 70%, put on a few drops of Histogel, mix to distribute the cells,
then cool the tube to harden the gel.  Once solidified, I'd remove the
pellet from the tube and breadloaf it like a piece of tissue, place in a
cassette and onto the processor (we started in 70% EtOH) for overnight
processing and embed as usual in the morning.  It always worked just fine.
What to you use for clearing?

Carrie Kyle-Byrne
Sr. Reseach Assoc., Pathology Lab
Lab Vision, Corp.
47791 Westinghouse Ave.
Fremont, CA     94539
ckbyrne <@t> labvision.com
www.labvision.com



-----Original Message-----
From: S Ladd [mailto:sladd <@t> hsc.usf.edu]
Sent: Tuesday, April 13, 2004 7:49 AM
To: m. van mkempen
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] histogel


Yes Yes Yes!!!!!!! If you don't fix the histogel in formalin before
processing it WILL DISSOLVE! I learned this the hard way! The instructions
say something vague like "process as usual". Of course, I informed Richard
Allen that many of us in research do not have formalin on our tissue
processors. Richard Allen said they were going to change the instructions.
Once you have fixed the histogel you can process it like you would any other
piece of tissue.
Sharron
University of South Florida

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of m. van
mkempen
Sent: Tuesday, April 13, 2004 10:18 AM
To: Histonetlist
Subject: [Histonet] histogel


Dear all,

I have a question concerning histogel!!
I'm working with fetal lung and I thought histogel would be a nice way
to handle then while embedding.
My question is do I have to fix the histogel before processing for
embedding in paraffine?
How is the normal procedure for tissues's in histogel??

Thank you for all your kind answers.

Regards,
Marta.

--
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*- mkempen -*
MAILTO:m.vankempen <@t> erasmusmc.nl
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