Dissotiation of cells from the bottom of a transwell - (Dec/06/2007 )

Hi everybody,

I was thinking of making migration invasion experiments more quantitative and easy, using 96 well transwell plates. In these kind of transwell I guess you cannot count migrated cells, but the best is to dissociate the cells that have passed the membrane, stain them somehow (calcinein etc.) and quantify them using a microplate fluorimeter. My question is? How would you dissociate the cells? I found commercially available solutions, is there also a simple home made solution?
Another question: once in the lower well cells need to be quantified: anybody has never tried just to add some Bradford reagent to quantify the amount of protein? that should be proportional to the number of cells. Maybe not sensitive enough?

thanks a lot for the help!

I dont think you would treat transwell anything differently from the normal ones. Many of your questions could be case-dependent and only your own test can tell.

Cant you just count the cell numbers directly? All other assays are indirect in nature and you have to calibrate with cell numbers, or it will only gives you a relative term. Cell counting is a bit more time-consuming, but its a direct method.