[Histonet] Microwave retrieval new question

[Flynn, Evelyn]

Dear Connie and others,
       In microwave antigen retrieval, I do the heating in three steps:
       1.  I heat a "water bath" filled with water for 5-10 minutes.  I  use a Rubbermaid plastic bowl, a 6 cup-
size.
       2.  In PLASTIC Coplin jars sitting in the water bath I heat the retrieval solution for 5-10 minutes.
 In order for the jars to sit in the water bath, I fill the slots with blank slides (otherwise the jar would float).
       3.  Carefully with forceps I remove the blank slides and put in the slides with the sections.  The solution
should come to the top of the jar to allow for evaporation.  Cover loosely to allow steam to escape.
I microwave the jars in the waterbath for 15-20 min.  Remove the jars from the waterbath, remove the covers,
and allow to cool for 20-30 min.
   
Good luck,
Evelyn Flynn


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of Connie McManus
Sent: Thu 10/21/2004 10:21 AM
To: 'Weems, Joyce'; 'marjorie lehman'; 'Amos Brooks'; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Microwave retrieval  new question
 
I find this all interesting as my pathologist has suggested that I try
the MW for our AR procedures.  We were thinking about using it for
proteinase K rather than using a citrate buffer or Tris.  I would assume
that the temp would be very important to control with the enzyme. So...
  For those of you who do MW AR, would you mind sharing your procedure?

Thnks bunches!!  *G*

Connie McManus
Utah Veterinary Diagnostics Laboratory
Utah State University
Logan, UT
Phone:  435/797-1891
fax: 435/797-2805
email: convmcm <@t> cc.usu.edu


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Weems,
Joyce
Sent: Thursday, October 21, 2004 5:15 AM
To: marjorie lehman; Amos Brooks; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Microwave retrieval

We were fortunate enough to have some left over capital dollars and have
purchased the new Biogenex Retreival Microwave. It is wonderful - saves
lots of time and gives consistency in staining! The only thing we would
change is to be have smaller containers for instances when we have just
a few slides to retreive. But we think its a winner for those of you are
looking to purchase it. 

Cheers, 
Joyce


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu]On Behalf Of marjorie
lehman
Sent: Thursday, October 21, 2004 7:19 AM
To: Amos Brooks; histonet <@t> lists.utsouthwestern.edu
Subject: RE: [Histonet] Microwave retrieval


Here is a Second Opinion!
I tried using a (Household) microwave for a long time and was not happy.
After
I got my Decloaking Chamber from Biocare I never looked back! 
Marge

-----Original Message-----
From:	Amos Brooks [SMTP:amosbrooks <@t> earthlink.net]
Sent:	Wednesday, October 20, 2004 8:00 PM
To:	histonet <@t> lists.utsouthwestern.edu
Subject:	[Histonet] Microwave retrieval

Ed,
         I strongly recommend that you don't try to put a coplin jar of 
retrieval solution with slides in it in the microwave and expect any 
consistant results. What happens with this method is you get a spike of 
heat then a temperature drop. Very often there is boil over and your
slides 
dry out. Ideally you should be looking for an even temperature for a 
defined length of time that can be well monitored.
         For this reason if you *must* use a microwave it should be a
lab 
grade oven that has consistant wattage and temperature control. It would

also be good to use one of the microwave pressure cookers to improve the

consistency. (I think Biocare Medical has them ... the guys that sell
Borg 
Decloaker).
         My ultimate recommendation is to stick to the tried and true 
waterbath or steamer method. It may take a bit longer but the
consistancy 
is better and there is far less variables in the procedure.
Just my opinion,
You know what they say 'bout opinions,
Amos Brooks

At 01:00 PM 10/20/2004, you wrote:
>Message: 10
>Date: Wed, 20 Oct 2004 11:30:26 -0400
>From: "Harris, Ed" <Hared <@t> FGHI.COM>
>Subject: [Histonet] Microwave retrieval
>To: "'histonet <@t> lists.utsouthwestern.edu'"
>         <histonet <@t> lists.utsouthwestern.edu>
>Message-ID: <BCBDA208D7498449A95432B874A9629752AAE1 <@t> fghmail.fghi.com>
>Content-Type: text/plain;       charset="iso-8859-1"
>
>Can anyone give me the pro's and con's of steamer/pressure (Decloaker)
vs.
>microwave retrieval ?
>Our Doc wants us to change from the Decloaker to microwave. Is a
household
>microwave adequate or do we need a lab grade unit?
>Does the microwave save time and improve turn around time?
>
>Thanks in advance
>Ed Harris  HT ASCP MT ABB



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