5 replies to this topic

[verena]

Hi!

I am looking for a procedure to stain collagen. What I have found so far does not really seem to fit my needs.
The key parameters and requirements of my setup are:

adherent fibroblast cell culture (not tissue slides)
staining the collagen secreted by the cultured cells
a procedure that does not require cell fixation (plus a non-toxic dye) for staining a living cell population would be highly welcome.

Thanks for any ideas and suggestions!

[verena]

no ideas at all?  
I am grateful for any advice!

Edited by verena, 11 October 2009 - 12:38 PM.

[bob1]

The living cell population thing is the difficult side of things - I don't know of any specific things against collagen other than antibodies, which you could try in a live environment, but probably wouldn't be very specific without the stringent washes.

[verena]

Thanks for your answer!
Ok, so I guess I will just have to fixate the cells...

Whats the standard method in this case?
What I mostly found was the Masson's trichrome staining, but I guess its only for histological samples, not for in vitro cell cultures.
So I think maybe antibodies are best?

[shyamadi]

I dont think masson trichrome has only been used for tissue sample staining. There are some reports using masson trichrome for adherent cell staining as well. We need to seed the cell in cover slip, followed by fixation as described for masson trichrome staining. Actually I was also kind of wondering, but finally decided to give a try.

[shyamadi]

I would appreciate if you share your experiences.