[Histonet] processing histogel

[Liz Chlipala]

Luciana

You process histogel samples essentially the same way you process
regular tissue samples.  I'm assuming you will be dealing with a about a
1 cm cube sample that you want embedded into the histogel and then you
would then slice the histogel cube (with the collagen construct) into 3
mm sections and process to paraffin.  You will need some type of mold to
make the histogel cube.  I would use a peel-away mold that is a bit
bigger than your construct.  If your construct is smaller than the 1cm
cube then you might be able to get away with a cryomold, or embedding
mole.  I would basically use the histogel as the manufacture states,
melt to the correct temp, pour a bit of it into the mold, place your
fixed construct on top of it and fill up the mold with more histogel,
let solidify and then place the entire sample in 10% NBF and then
process to paraffin.  We have dealt with a lot of different constructs
and sometimes you just have work with them a bit to get what you need
out of the paraffin sections.  I would not waste your cultured samples
right away but work out your technique on the construct alone initially.

Liz

Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Manager
Premier Laboratory, LLC
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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Luciana
Bastos
Sent: Monday, February 14, 2011 6:06 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] processing histogel



Hello,
We are working with cell culture embedded into three dimensional (3D) of
collagen gel type I.  The samples were fixed in 10% formalin for 24 h.
Even after fixation, the gel used in our preparation was too soft to be
embedded directly in paraffin. To circumvent this problem, we wold like
to dehydrated and embedded the samples in Histogel (Perk Scientific
Inc., Devon, PA, USA).11 V.M. Freitas and R.G. Jaeger, The effect of
laminin and its peptide SIKVAV on a human salivary gland adenoid cystic
carcinoma cell line, Virchows Arch 441 (6) (2002), pp. 569-576. Full
Text via CrossRef | View Record in Scopus | Cited By in Scopus (12)11
However, we need some help to how dehydrated the collagen gel and the
histogel (solutions, concetration, time, ect.) toparaffin-embedded and
stained by haematoxylin-eosin (H&E).
Thank you,
Luciana


 
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