Boyden chamber - isolation of cells? - (Oct/30/2008 )

Hello, can someone tell me if it is possible, using the Boyden's chamber, to isolate the migrating cells and use them for RNA isolation? Normally, what people are doing - they are using the Boyden's chamber method to investigate different chemotactic agents, all the articles I read are fixing the cells or staining them for subsequent countings. But I was wondering if you can isolate the cells and use them, for example, for RNA isolation? Have you heard of someone doing this?

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I haven't heard of it - but it seems plausible.
When you open up the chamber you can literally scrape the cells off one side of the membrane.... I don't know if you would have enough cells for isolation though (maybe with amplification).

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Thanks for the reply Aaron. In fact, I haven't seen a Boyden chamber (only read about it), so it's a bit hard for me to imagine how exactly it works without seeing the procedure. I know it's a porous membrane and the cells migrate on the other side of the membrane. So you say that it's absolutely possible to scratch the side of the membrane with the non-migrating cells, so that on the membrane only the migrating cells will remain. There should be a soultion which can detach the cells from the membrane or to add the lysing solution directly to the membrane (in case there are no more cell left on the other side), but I agree that the number of cells will probably be a problem...

Does somebody has a better idea to isolate migrating from non-migrating cells for RNA isolation?

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I'd say it's possible, but not easy, yeah try and get your hands on one - or find a nearby lab doing them. The membranes about the size of a credit card - you can tear it - but if you are careful you shouldn't.
You could also try buying a transwell - the migrating cells migrate into a lower chamber - you could then isolate them easier.

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