[Histonet] Immuno Bed Plastic Question

[Robert Schoonhoven]

Well..... If it is truly GMA then you will not be able to remove it.  What you describe is one of several methods to remove MMA.   You will also lose the occasional section on exposure to the higher ( 70% +) grades of alcohol.
 
If it is GMA I'd suggest you place your slides directly in PBS and then start your IHC procedure.

Robert Schoonhoven, HT/HTL (ASCP)


________________________________
From: Allyse Mazzarelli <allyse124 <@t> gmail.com>
To: histonet <@t> lists.utsouthwestern.edu 
Sent: Wednesday, February 13, 2013 1:19 PM
Subject: [Histonet] Immuno Bed Plastic Question

Hi histonetters!

I have posted on this forum a while ago asking for opinions regardnig new
types of plastics that work well with IHC. It was brought to my attention
that "Immuno-Bed" (Polysciences) works rather well. I recently purchased
the plastic, infiltrated/embedded well, and sectioned great, BUT, when I
removed the excess GMA/rehydrated and labeled with my antibodies, NOTHING
WORKED! Nothing showed up! Only DAPI!! The mechanism I used for removing
GMA and rehydrating were as follows:

1.) 3 changes of 30 minutes each in 60 Celsius xylene. (Note: I recently
let the slides sit in 60 Celsius xylene overnight too... but it did not
work!!)
2.) 3 changes of 10 minutes each in 100% EtOH.
3.) 15 minutes in 95%, 75% & 50% EtOH
4.) 5 minutes in water
5.) 3 changes of 5 minutes each, PBS.

I continued to block and apply my antibodies as normal. (Antibodies are all
brand new, not duds!)

Since this plastic was formulated specifically for IHC, I am at a loss. If
anyone has used this plastic before, any insight directed my way would be
extremely helpful!!

Thanks,

Allyse Mazzarelli
Histologist
NEUROTECH USA INC.
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