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how to make nonadhensive cells attach to coverslips without affecting cellular s


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#1 shm267

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Posted 19 November 2009 - 02:50 AM

hello everyone, I encountered an awkard situation. I have two type of cells, one can tightly adhere to the fibronectin coated coverslips, and the other very weakly. we want to examine the f-actin and other related proteins by phalloidin staining or IF, which requires that the cells should stay on the coverslips whether they can adhere to the matrix or not. however, the nonadhensive cells often washed off when carrying out these staining protocols. of course, they say you can air the cells before fixation, but I am wondering if this treatment can damage the cell morphology and structure. I am very appreciated if anyone can give me some suggestions, thanks a lot.

#2 bob1

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Posted 19 November 2009 - 04:20 PM

It's kind of tricky for weakly adherent cells, the most you can do is coat the coverslips with something, as you have already. There are a range of ways of fixing cells/tissues to slides during the fixation process, most of which involve air-drying the sample. These processes do dehydrate the cells and change morphology a bit.

I guess you could try making a cell clot - basically making a lump of cells by centrifugation and then fixing them and sectioning as you would for normal histology.




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