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collagen staining in cell culture


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5 replies to this topic

#1 verena

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Posted 09 October 2009 - 12:16 AM

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!

#2 verena

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Posted 11 October 2009 - 12:38 PM

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

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


#3 bob1

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Posted 11 October 2009 - 03:53 PM

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.

#4 verena

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Posted 15 October 2009 - 10:32 AM

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?

#5 shyamadi

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Posted 21 January 2013 - 05:52 PM

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.

#6 shyamadi

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Posted 21 January 2013 - 05:54 PM

I would appreciate if you share your experiences.




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