Two FISHy questions

[simon walker (BI)]

Dear Listers,

I have two questions relating to FISH.  Firstly, does anyone have experience
of doing FISH in (mouse) oocytes?  We are looking to prepare cells for
confocal microscopy and need to find a reliable protocol to rapidly fix these
cells, with subsequent staining and mounting to preserve 3D structure.

Secondly, we are doing the same FISH staining in ovary sections, but due to
ab/probe penetration are limited to a section thickness that is less than a
complete oocyte.  We are hoping to image serial sections and reconstitute the
sections into a 3D volume.  Does anyone know of software which will allow
you to easily align images taken from different tissue sections?  Imaris has
something called Autoalign, which seems to be what I'm after, but I was
rather hoping to find something I didn't have to pay for..

Thanks in advance,
Simon

[Calvo-Bado, Leonides]

Hi,

I am looking for protocol to embed fixed skin tissue either with resin or parafin for FISH. Also, if there are companies that I can send samples for microtome sectioning. I have done some embedding using Technovick resin and sectioning but I need to speed up my work and I would prefer to pay for the sectioning. Any help or advice, I would really appreciate it.

Thanks

Leo

 

Dr. Leo Calvo-Bado

Postdoctoral Research Fellow

Environmental Microbiology Group

Department of Biological Sciences

The University of Warwick

Coventry CV4 7AL

United Kingdom

 

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