Amy Self <ASelf@gmhsc.com> wrote:

>	Is there another stain other than alcian blue pH 2.5 that can be
>used to stain GI biopsy when looking for goblet cells.   Someone told me
>that there is a stain that came from Allegiance that was just a quick dip
>and that was all there was to it.  I have heard of a diff quik stain from
>Allegiance but I thought that it was used for H. pylori.  Thanks
>

The short answer is that any basic (cationic) dye at pH 2.5 to 3
will stain the same things as alcian blue at pH 2.5. There are
two differences, however: (1) Cell nuclei are stained as well as
the mucus, and (2) Other basic dyes are not changed to insoluble
pigments after binding to mucus, so you need to be careful not to
lose the stain during washing & dehydration. 0.5% toluidine blue
is an excellent general-purpose basic dye.

If the goblet cells of interest contain sulphated mucus they can be
stained selectively (without nuclei) by a basic dye at pH 1.
Another selective method and simple method is that of I.D.Heath
(1962) - Quart J Microsc Sci 103: 457-475. Sections are stained for
5 to 30 minutes in a 0.03% solution of a basic dye (toluidine blue
or neutral red) in 5% aluminium sulphate. Wash in water, rinse for
a few seconds in 70% alcohol to remove any loosely bound dye from
stuff other than sulphated carbohydrates (mucus, cartilage matrix,
mast cell granules), complete the dehydration in 3 changes of 100%
alcohol, clear and cover. The aluminium salt provides an acid pH
and also excludes dye from nuclei. Heath's publication is well
worth reading, and his method is easy and specific. Unfortunately
Heath wrote of the virtues of "nuclear fast red" in this technique. 
Obviously he didn't mean C.I.60760, but people might
have used it and concluded that the method was no good. (It might
be feasible to use real N.F.R. as a nuclear counterstain; I haven't
tried that.)

---------------------------------------
John A. Kiernan,
Department of Anatomy and Cell Biology,
The University of Western Ontario,
London,  Canada  N6A 5C1
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