Re: Immunostaining in Neuropathology

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From:Amos Brooks <>
To:"Timothy R. Wheelock" <>, histonet <>
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    I think the best way to determine how feasible adding
immunohistochemistry (IHC) to your repertoire is to find out how many
specimens you send out to other labs for it. Then find out how much you pay
for such service.
    Then find a vendor, preferably one with a wide variety of antibodies in
their catalog including the ones you send out for the most. Once you have
chosen a vendor contact a sales rep for the company. Find out the start up
costs. This would include buffers, staining racks, detection kits,
chromagens, pipettes (if you dont have them already) etc. Some companies
will offer samples of these products to try out. Then compare the costs
(keep in mind long term) of starting up your own vs. sending the work out.
    I would also highly recommend that you review the protocols you want to
use and make sure you fully understand them. Don't try to invest all this
money with out being sure your not getting in over your head. A fantastic
IHC reference that should be in all IHC labs just because of its simplicity
is a book DAKO has I can't remember the title. It is an 8x10 paperback book
explaining the theory of IHC staining and tips for troubleshooting staining
problems, it's about 100 pgs long and a great reference.
    If it turns out that starting up IHC service is not feasible most
reference labs prefer if you send them a block to work with but usually will
accept the unstained slides. (thats how we do it ... although we are not
specialized to neuro)
good luck you are in for an adventure
Amos Brooks

"Timothy R. Wheelock" wrote:

>         I manage and single-handedly work a neuropathology lab at a
> Brain Bank, and I am considering adding an immuno-staining capability to
> my classical staining routine. I have some general questions to ask
> before making such a move.
>     (1) Has immuno-staining become a  standard routine practice in
> neuropathology these days?
>           We concentrate on degenerative and psychiatric disorders, and
> rarely see tumors, so we don't  have
>             to worry about tumor identification. Or is it only used
> occasionally, for example,  to distinguish Diffuse
>             Lewy Body disease from Pick's disease using Alpha B
> crystalline antibodies? I get different answers
>             from different pathologists.
>     (2) If it is used routinely, what sort of immuno-stains are used?
>                 From one pathologist I get a list of Tau, Ubiquitin,
> Amyloid, and Alpha B crystalline. From another
>                 I'm told that he rarely uses immuno-staining at all
> except occasionally and that my Bielschowsky
>                 stains and LHE's show more than 99% of what's needed to
> make a complete and detailed
>                 diagnosis.
>     (3) Whether routine, frequent or occasional, does the neuropathology
> lab tech do the immuno themselves, or simply out-source the immuno work
> ?
>             One pathologist says that his histo-tech sends the unstained
> slides to another lab that specializes in
>             immuno and gets the slides back in one day,  rather than his
> tech having to re-invent the wheel, at
>             considerable expense and time, in adding an immuno routine
> to her schedule.
>     (4) If the immuno function is out-sourced what sort of fees can we
> be expected to encounter for such a
>           service?
>     (5) Is there a review paper or protocols book that I could consult
> that reviews the immuno-stains used in neuropath (other than tumors)?
>         Thank you very much for any information and advice that you
> could give me,
>          Tim Wheelock

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