[Histonet] get back eosin stain?

[Yak-Nam Wang]

Do you know why this is? In the past, when I have cultured cells and frozen
without fixation the cytoplasm stains pink. Is the lack of pink in these
cells due to the fixation? Do you know if there is a way of fixing this (no
pun intended!).

Thank you
Yak-Nam


On 8/18/08 7:22 AM, "Andrea Grantham" <algranth <@t> u.arizona.edu> wrote:

> 
>> 
> 
> I see this a lot with cultured cells - even when processed and
> embedded in paraffin.
> 
> Andi Grantham
> 
> 
>> 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)       :
> :...................................................................:
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> 
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