My tissue is a mice skin tissue and, liver, and kidney.
>To: Thu-Thao Pham
>CC: Abigail.Mackey@ul.ie, HistoNet@pathology.swmed.edu
>Subject: Re: Pretreatment.
>Date: Mon, 10 Dec 2001 13:24:44 -0700
>what is your tissue? if it is bone or cartilage HIER can be a real problem
>Thu-Thao Pham wrote:
> > Hi everyone,
> > As some of you may know that I post a questions regarding CD31 and
> > pretreated the slides with DAKO Target Retrieval Solution. However, my
> > tissue got eaten up and the mophology look back. Some tissues even about
> > come off the slides. I try to be very careful when I take the slides out
> > washing the slides.
> > I did as followed:
> > 1) Deparaffinized (Histoclear to water)
> > 2) Washed slides in TBST 3x for 5 min.
> > 3) Pretreatment with TRS by steam for 20 min.)
> > - first heat the TRS to boil and let the TRS
> > solution become clear before place slides into
> > the TRS heated solution (95C).
> > 4) Remove the jar from steamer and let it cool to room temperature for
> > 30min.
> > 5) Gently remove the slides and washed in ddH2O 3x for 5 min.
> > 6) Washed slides in TBST 3x for 5 min.
> > 7) Proceed with 3% H2O2 for 5min.
> > 8) Washed TBST
> > 9) CD31 diluted in 1% BSA (in TBST).
> > 10) Followed by DAKO kit
> > After preteated my slides can I leave at room temperature for at least
> > hour to cool down?
> > Many thanks if anyone can help.
> > Thao
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