Imaging of single cells in organotypic slice cultures

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Sher
Sher's picture
Imaging of single cells in organotypic slice cultures

Hello,

Could anyone give me any advice on how I might try to label and image single cells in-vitro in organotypic slice cultures? I have never done experiments like this before so don't know where to start! I can tell the cells I'd need to label by morphology but don't know how to target them with for example GFP. I would greatly appreciate any advice anyone could give!

Thanks!

Guy Sovak
Guy Sovak's picture
Hi there are couple of

Hi there are couple of protocols, as I dont know what type of tissue you are using I found one from rat brain:
http://www.natureprotocols.com/2006/12/29/staining_protocol_for_organoty.php
You can use our customized search engine and you will find more.

jentennis
jentennis's picture
If you only want to fill a

If you only want to fill a single cell, you can do intracellular fills using lucifer yellow which can later be converted into a permanent record using immunoperoxidase staining. Here is a paper that might help you get started:

Intracellular injection in fixed slices: obtaining complete dendritic arbors of large cells
Journal of Neuroscience Methods, Volume 119, Issue 1, 15 September 2002, Pages 23-30
Christopher J. Pace, David G. Tieman, Suzannah Bliss Tieman