Sphere staining protocol (agarose)
Posted 15 February 2012 - 10:16 AM
I am growing putative stem cell spheres and I would like to use them for immunofluorescence.
I have seen many different ways of doing it. Some just put the sphere onto the slide, others include the spheres in agarose and them either freeze it and ultra cut or include them in parafin and then cut them.
Is anyone staining spheres that would be kind enough to share somne thoughts.
What technique should I use?
Posted 15 February 2012 - 10:48 AM
Embedding the spheroids in agarose is the better option if you don't use confocal, but obviously it takes rather longer.
Edited by Tabaluga, 15 February 2012 - 10:49 AM.
Posted 16 February 2012 - 02:39 AM
Thanks for the help.
Do you have the protocol for that? I am starting this work int he lab so no one has an idea on how to do it.
Can I kindly ask you for your advice?
Posted 16 February 2012 - 04:29 AM
Google Books => "Methods in endothelial cell biology" by Hellmut G. Augustin => starting from page 115
They do it with endothelial cell spheroids, but the basic protocol should be applicable to stem cell spheres too. It's with a collagen gel, though.