in situ hybe

M. Hertel hippocampus at
Tue Jan 30 14:16:53 EST 2001

Hello Rachel
The film emulsion should stay on the slide! it should be liquid enough
to adhere in one thin layer..( dip the slide to the emulsion in one
single smooth movement if you stop, or do it more than once you will get
unevenness to the layer. Prepare some example slides in a extra box, you
can develop them after the minimum exposure time to see if the important
samples can stay longer or if background is coming up)
All the counter stain (H/E e.g.) is done through this layer without any
In the end after you embedded Your sections with a cover slip You can
rub off the film emulsion on the back of the slide for better
microscopy, I do it with the rough side of a kitchen sponge but be
careful that the glass is not scratched...
good luck!!

Rachel wrote:
> I am performing a radioactive in situ hybridization study and have come
> to the point of developing my slides. Two tester slides have me quite
> confused. Is the NTB-2 emulsion supposed to stay on the slide,
> effectively covering the sections, or is it supposed to come off to
> reveal the silver grains? All protocols I have seen neglect to mention
> this detail, and I have one slide with a emulsion layer which won't come
> off and one whose emulsion layer is coming off in pieces.
> Thanks for your advice.
> Rachel Geddy
> McGill University

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