RE: [Histonet] manual IHC queries

From:Dana Settembre



Louise,
I used to always use "Parafilm" and I sometimes still do.
I cut it up into sizes that I need.
It works marvelously and is cheap.
 

Dana Settembre, HT ASCP
Immunohistochemistry Lab
UMDNJ - University Hospital
Newark, NJ    USA


>>> "Tarango, Mark"  12/12/07 11:53 AM >>>
I do sometimes use coverslips when doing IHC by hand.  The humidity in
Las Vegas is pretty low, so evaporation is always an issue.  The
Ventana's instruments use a coat of oil, over the aqueous antibody
solution, to prevent evaporation.  You could try using a lightweight
mineral oil too.

Mark Adam Tarango HT(ASCP)
Histology & IHC Supervisor
Nevada Cancer Institute
One Breakthrough Way
Las Vegas, NV  89135
Direct Line (702) 822-5112
Treo (702) 759-9229
Fax (702) 939-7663
  

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu 
[mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of louise
renton
Sent: Wednesday, December 12, 2007 12:36 AM
To: Histonet@lists.utsouthwestern.edu 
Subject: [Histonet] manual IHC queries

Hi all,

Firstly: As a matter of interest - when using small amounts of primary
antibody does anyone use  coverslips to spread the solution over the
section
and keep it from evaporating during the incubation? (i do use a
humidity
chamber also).  I have been doing this for ages and wondering if there
is an
alternative to use other than the coverslips that may be cheaper - any
suggestions??

secondly, Although 30min incubations are recommended, is there any
value
to
increasing incubation times of secondary and ABC complexes to improve
sensitivity? In other words is there a maximal binding period after
which no
improvement will be noted?

I hope I have made myself clear here....these are just passing
wednesday
thoughts in preparation for the usual  friday meltdown!
-- 
Louise Renton
Bone Research Unit
University of the Witwatersrand
Johannesburg
South Africa
"There are nights when the wolves are silent and only the moon howls".
George Carlin
No trees were killed in the sending of this message.
However, many electrons were terribly inconvenienced.
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