Re: Pretreatment.

From:Amos Brooks

    I'll bet it is your slides. Charged slides are a must. Also it would be
a good idea to keep a close eye on the temperature of the retrieval
solution. If the slides are actually boiling in the solution, it is too hot
and is unnecessarily agitating the slides. I would also caution against
using float bath adhesives being used in conjunction with charged slides.
The archives have countless explanations of  why this goes wrong. Beyond
that I'd look into the processing of the specimen.
Good luck,
Amos Brooks
----- Original Message -----
From: "Thu-Thao Pham" 
To: ; 
Sent: Monday, December 10, 2001 1:10 PM
Subject: Pretreatment.

> 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 an
> hour to cool down?
> Many thanks if anyone can help.
> Thao
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