Re: Fixation times for Immuno methods.

<< Previous Message | Next Message >>
From:Geoff McAuliffe <mcauliff@UMDNJ.EDU>
To:Victoria Baker <>
Content-Type:text/plain; charset=us-ascii

Victoria Baker wrote:

> Hi Everyone!
> The controversy over fixation and IHC has reared its'
> ugly little head again.
> So rather than bang my head against the wall from
> frustration, I thought I'd put it out and see what
> others are doing.
> The tissue is either rat or mouse, the size of the
> tissue is usually 5-6mm in diameter and approximately
> 4mm in depth.  This is a research facility where time
> isn't always critical (except when something is needed
> yesterday).
> So if anyone is working with this under similar
> circumstances do you think you could take a minute and
> just answer a couple of questions?
> 1.  How long do you fix your tissues for? 10 hours,
> >10 hours, 12 to 24 hours.

Depends on the antigen I am interested in. For some antigens 2 hours is
the max., others overnight is fine.

> 2.  Do you do fixation on or off of your processor?

Don't have a processor.

> 3.  Do you use different fixatives for different
> procedures?

Different fixatives for different antigens. Everything depends on the

> 4.  How long is your overall processing time and what
> do you use to hold tissue in after fixation?

Hold the tissue in phosphate buffer or sucrose. Processing time varies
with the antigen in question.

> 5.  This one may cause some controversy.....Does
> anyone cap their blocks after they have finished with
> them for storage.

I cap with melted wax, old school.

> 6.  Does anyone use a xylene substitute and which one
> do you use.

I use xylene which I have an enormous surplus of.  Poor results with the
orange-odor products.

    In case I forgot to mention it, everything depends on the antigen
you are looking for.

Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029

<< Previous Message | Next Message >>