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Collagen formation by fibroblast

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Collagen formation by fibroblast

Postby Yeditepe » Thu Oct 06, 2011 4:17 pm

I want to observe and take good pictures of collagen produced by fibroblast (L929) under the microscope. Can anyone help me with the protocol.

what can I do to stain collagen and observe them?

Thank you...
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Postby biohazard » Fri Oct 07, 2011 6:33 am

I'm not an expert in this field, but I'd guess you could do it so that you grow your cells under conditions where they form collagen and then take some cells at regular intervals (depending on the production speed you could take, say, one sample every two hours for 24 hours, or whatever that seems useful) and stain them with some general antibodies specific for common cell surface markers of fibroblasts and with a nucleic stain (to visualize the cells), as wells as with collagen-specific abs. That way you could see the process under a confocal microscope and take an image at each time point.

Watching the images in a chronological order would hopefully provide a kind of animation of collagen production.
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