Re: Pretreatment.

From:Dana Settembre

Hello Thao,
In addition to all the suggestions you received on your sections coming off or threatening to come off, 
I suggest that when you cut your sections on Superfrost Plus slides, that you air dry them completely to make sure all the water is dry or gone then bake them in a 60 degree oven from 30 to 60 minutes.  I have tough times with skin too.
Dana Settembre
University Hospital- UMDNJ, Newark, NJ

>>> Thu-Thao Pham  12/10/01 03:39PM >>>
My tissue is a mice skin tissue and, liver, and kidney.

Thao


>From: rueggp 
>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 
>in
>my experience.
>patsy
>
>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 
>to
> > come off the slides. I try to be very careful when I take the slides out 
>and
> > 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 
>an
> > hour to cool down?
> >
> > Many thanks if anyone can help.
> > Thao
> >
> > _________________________________________________________________
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