[Histonet] help!!

[Caroline Miller]

Blanca,
Here are my feelings on this, and I am sure a lot of other folks have feels
here too, so please chime in.

1 - I feel that most clinical labs are more on the IHC bandwagon and
research labs are IF (with the exception of IgG staining in kidney biopsies
or bullous disease in skin- which is because the antibodies don't like
formalin fixing (if this is now wrong I am sorry, I haven't been in a
clinical lab in quite a while). Research labs are often also working with
genetically encoded fluorophores such as GFP, YFP, mCherry
2 - Formalin fixation (especially over fixation) can often lead to a large
amount of autofluorescence in the 488 region, which is a common place for
secondary antibodies and also GFP. Research labs have a lot more control
over their fixation protocols.
3 - The microscopes commonly available to clinical labs are bright field
scopes and in research labs fluorescent scopes
4 - Fluorescence can provide more contrast to a positively localized
fluorophore, but sometimes at the detriment of viewing the overall
morphology of the tissue like you get with bright field IHC and a nuclear
counterstain.
5 - Research lab protocols are often very 'experimental' and can lead to
increased tissue damage, which again is not viewed under the fluorescence
microscope (as much). Clinical labs have lots of experience and also
defined protocols that work well in the IHC / bright field space.
6 - the only real difference is the detection method, you can use any
primary antibody with either ABC/ impress / enzyme based methods or with
fluorophore conjugated secondaries.

So, in short - no *real* reason, but mainly that is the way things shook
out.

I could go on about researchers not understanding how to take photos on a
bright field scopes too, but that is too broad a statement, but as a core
director I saw them being more comfortable with the fluorescent methods :)

mills




On Thu, Apr 13, 2017 at 6:09 AM, Blanca Lopez via Histonet <
histonet at lists.utsouthwestern.edu> wrote:

> Hello!
> I just need a help with a simple question...Is anyone can explain me what
> is the purpose between performing immunohistochemistry and
> Immunofluorescence?
> Thanks  :)
>
> Blanca Lopez
> Histotech (ASCP)
> UTSW Tissue Resource K1.210
> Simmons Comprehensive Cancer Center
> UT Southwestern Medical Center
> Telephone: 214-648-7598
> Email: Blanca.Lopez at utsouthwestern.edu
>
>
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-- 
Caroline Miller (mills)
Director of Histology
3Scan.com
415 2187297