[Histonet] get back eosin stain?

[Mark Tarango]

Were the cells that you cultured this time of the same type as before?
 Is it the same media?  Were they cultured for just as long?  I still
say try the PAS, you originally asked for the "look" of an H&E.

I did spit on one once and the stain came back a little.  Sometimes I
do strange things like that.  Probably shouldn't have even mentioned
it.

Mark


On Mon, Aug 18, 2008 at 8:33 AM, Yak-Nam Wang <ynwang <@t> u.washington.edu> wrote:
> 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.:
>>>
>>>
>>>
>>> _______________________________________________
>>> Histonet mailing list
>>> Histonet <@t> lists.utsouthwestern.edu
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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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>
>
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