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.:
>>>>
>>>>
>>>>
>>>> _______________________________________________
>>>> Histonet mailing list
>>>> Histonet@lists.utsouthwestern.edu
>>>> http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>>>
>>> .....................................................................
>>> : 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@u.arizona.edu) :
>>> :...................................................................:
>>> http://www.cba.arizona.edu/histology-lab.html
>>>
>>>
>>> _______________________________________________
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>>
>>
>>
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>
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