[Histonet] get back eosin stain?

[Bryan Llewellyn]

It is sometimes possible to stain tissues with eosinol when they do not 
stain with eosin Y.  Eosinol is the free anion of eosin Y and is not soluble 
in water.  You can sometimes buy it, or can make it from a strong aqueous 
solution of eosin Y by dropping in concentrated hydrochloric acid until 
maximum precipitate is obtained, then filtering and washing several times 
with distilled water to remove the acid.  When dried, a small amount added 
to the final xylene (yes, xylene) will colour the tissue.  When dark enough, 
coverslip.

Bryan Llewellyn


>>>
>>>> Hello,
>>>>
>>>> I have a colleague with a staining problem. He has cells grown on 
>>>> polymeric
>>>> scaffolds. He can not embedd in paraffin due to a mismatch in the 
>>>> processing
>>>> and his scaffold material; he has thus embedded in OCT.
>>>>
>>>> This is his procedure:
>>>> Fix cell seeded scaffold in formalin (these are delicate samples
>>>> thus, fixing before embedding)
>>>> Cryoprotect in grade sucrose to 30% sucrose
>>>> Embed in OCT
>>>> Section at 6um
>>>> General H&E staining-using 1 min for Hematox and 1.5 min for Eosin 
>>>> (following
>>>> one of Richard Allan suggested protocols).
>>>>
>>>> Problem: there is no eosin staining. The cells are just purple,
>>>> there is no pink at all. We are not sure why this is. Is this caused
>>>> by fixation?
>>>>
>>>> Does someone have any suggestions on how he can get the classic H&E
>>>> stain look for these samples? He has tried fresh solutions as well
>>>> as running other tissue (unfixed) through. It doesn't seem to be the
>>>> solutions used.
>>>>
>>>> Any suggestions would be appreciated. Thank you
>>>> Yak-Nam
>>>>
>>>>
>>>> Reserach Associate
>>>> CIMU
>>>> University of Washington
>>>> Box 355640
>>>> Seattle, WA 98195
>>>>
>>>> Tel.:
>>>>
>>>>
>>>>
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>>>
>>> .....................................................................
>>> : Andrea Grantham, HT(ASCP)     Dept. of Cell Biology & Anatomy     :
>>> : Sr. Research Specialist       University of Arizona               :
>>> : (office:  AHSC 4212)          P.O. Box 245044                     :
>>> : (voice:  520-626-4415)        Tucson, AZ  85724-5044    USA       :
>>> : (FAX:  520-626-2097)          (email:  algranth <@t> u.arizona.edu)       :
>>> :...................................................................:
>>>            http://www.cba.arizona.edu/histology-lab.html
>>>
>>>
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