Re: coverslips for In-situ staining
You don't need any special kind of coverslips. We are
routinely doing ISH with standard glass coverslips, and
it's working perfectly.
Briefly:
- Apply hybridization solution (100 ul/slide) on
dehydrated and air-dried slides
- Place the (ethanol-rinsed and air-dried) coverslips
on the slides, avoiding air bubbles
- Place the slides in a preheated oven (we are using an
oven with a removable metal tray) for denaturation
(we are doing genomic in situ hybridization to tissues,
so the denaturation is pretty harsh - 85 C, 8 min)
- Place the slides in airtight containers, with a bit
of tissue paper moistened with base constituents of
the hyb buffer
- Incubate
We are using a standard home-made hybridization buffer
(containing formamide, SSPE, Denhardt's, and ssDNA). No
problems with drying, with hybridization times up to two days. There
is a noticeable amount of vapors in the oven when we open
it, so some of the buffer is evaporating, but not too much.
Probably a good idea to avoid extensive exposure to the
formamide-containing vapors, though.
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Mikael Niku URL: www.helsinki.fi/~mniku/
University of Helsinki Dept. Basic Veterinary Sciences
- Mitäkö mieltä olen länsimaisesta sivistyksestä?
Minusta se olisi erinomainen ajatus!
- Gandhi
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----- Original Message -----
From: "Garza-Williams, Sara"
To:
Sent: Thursday, October 18, 2001 1:27 AM
Subject: coverslips for In-situ staining
> Can someone tell me where I can purchase coverslips for doing an In-situ
> assay. They are the type that you use when your doing the denature or
> hybridization part of the assay. They are plastic/glass with a type of
> rubber "gasket" which allows you to add the probe and incubate the slides
> without the probe drying out. Thanks in advance!
>
> Sara A. Williams
> Anatomic Pathology Supervisor
> The Children's Hospital
> Denver, CO
> 303 861-6177
>
>
>
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