I have developed a T cell assay using 48well culture plates. However I now want to develop the assay further by separating cell populations using transwells/thincerts in the same assay. So far I have only found that transwells/thincerts are made for 24well plates so I have done a couple of experiments using the larger sized wells (just by doubling the number of cells and volumes of reagents in each well) however the results I am getting are completely different from the previous experiments. Is doubling incorrect? should I be increasing the # of cells and volumes significantly more in view of increased surface area? or what would be much easier is if I could find transwells for 48 well plate. Any advice much appreciated.
I had the same kind of problem with my virus infections. When I wanted to shift from 96-well plates to 24-well plates, the infection rate was quite different though I just increased the volume without changing the virus titer or cell density. I think it is due to the dynamics of the components such as ions and small particles in the solution, surface area, the way of heat transfer and also gas exchange from the surroundings. I think you can try different volumes, cell density and concentration of other stuff to overcome the changes. I hope that works. Goodluck.
In order to maintain the same density of cells you need to calculate cell number per cm2 of growth surface area that you were using in the 48-well plates and use that same density with the 24-well inserts.
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