Re: [Histonet] stain for tight junctions

From:Geoff McAuliffe

Hi Kathy:

    You can put peroxidase in the vascular system and watch it leak, or 
not leak, out of the capillaries. I suppose you could put it in the 
lumen of the G-I or UG tract as well, if that is the tissue you are 
interested in. I do not know of the specific details of this method, but 
it has been used since the 1970's,  and probably earlier, to show 
transcapillary transport.  Do a google search for "peroxidase tracing" 
and see what you come up with.
    Have a look at J. Neurocytology 10:607-643, 1981 for a method using 
Evans blue as a tracer. It is a small molecule and crosses most 
endothelia easlily but it might be what you want.
    I seem to remember that lanthanum has also been used to trace 
leakage, or lack thereof, across epithelia.
    Keep in mind that these methods are quite "old" and, while 
well-documented in the literature, may not show up in an internet 
search. You may have to go to the library.


Walters, Katherine S wrote:

>Hi all,
>I have kind of an interesting problem.  We want to look at tight
>junctions in cells.  We need a small molecule stain that will not be
>taken up by the cells.  We want the stain to get between the cells and
>be stopped by the tight junctions.  We do not want to use EM and we do
>not want to use fluorescence.  Has anybody done this, or have an idea
>about how to do this?
>Thank you,
>Kathy Walters
>Central Microscopy Research Facility
>University of Iowa
>Histonet mailing list

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

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