[Histonet] get back eosin stain?

[Yak-Nam Wang]

Thank you for all your suggestions.
He tried extending the eoson staining time and this improved things (there
is at leasat some pink now). Next he will try eosinol.

Thanks again
Yak-Nam


On 8/18/08 3:07 PM, "Bryan Llewellyn" <llewllew <@t> shaw.ca> wrote:

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