Re: Need Antigen Retrieval Help

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From:Amos Brooks <> (by way of histonet)
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    This is the main reason we do not use microwaves for IHC procedures
The microwaves vary in power and unless you want to spend a fortune on a
microwave with all the sensor attachments.
    If this is a clinical setting, I would redo the run. You dont want to delay
resulting, there is an anxious patient on the other end of those slides. If its
not clinical then I'd let it go just out of curiosity.
    On another almost related topic: when I was a student, the hospital I was
finishing my training in, microwaved large buckets of formalin with with
lids on
containing breast specimens. This was to accelerate fixation. Anyway, I didn't
know that there was anything in the microwave and I opened the door. The
lid had
popped off while the solution heated. By the way the microwave was not under a
hood. So the door pops open and I peek in. A cloud of gaseous formalin belched
out of the oven and went right into my eyes and nose. I recovered soon, but an
apprehension toward microwaves continues to this day.

"Bennett, Catherine (Katie)" wrote:

> Hi all!
> I'm starting a double staining IHC run this morning that is intending to
> take two days, and I may have already committed a fatal error on step one,
> so I'm wondering if I should continue or start over.
> I am doing microwave antigen retrieval and had a problem with the solution
> boiling over and completely out of the coplan jar.  My protocol calls for
> microvaving slides in a coplan jar of antigen retrieval for 2x 5 min, but
> after just 1.5 min all the solution is boiled over and gone.  Needless to
> say, I didn't catch it in time and the slides may have gone through some
> microwaving without any solution in the jar.  I had placed the coplan jar in
> a small dish of shallow water, so the microwave was very humid at least.
> I'm guessing the microwave is much more powerful than the one used in the
> protocol I am using and I should experiment with which power setting I
> should use.  However, in the meantime, are the slides from today toast
> because they have been "cooked" without being in the antigen retrieval
> solution, or can I continue with them?
> Any suggestions from others on preventing boil-over would also be much
> appreciated.
> *********************************
> Catherine "Katie" Bresee Bennett
> Sr. Technical Associate
> Lovelace Respiratory Research Institute
> Albuquerque, New Mexico

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