I've been having trouble with this assay. In the control transwell chambers I see that the cells migrate uniformly, but in the ones with matrigel, only the cells in the edges invade (there is a circle without cells in the middle).
I'm using the Corning® BioCoat™ Matrigel® Invasion Chambers, so the problem is not due to the coating of the matrigel.
Hi Sofia,
I have heard people complaining about this issue, some of them say that cells that migrate in the border of the insert should not be considered and only the cells in the middle should be counted. However, seems like in your case you barely have cell migrating in the middle of the insert, so I wonder if it’s a matter of the volume of Matrigel that you plate. It’s a silly thing but If the amount of Matrigel used is small, it can solidify as a “drop” of liquid in a surface and don´t touch the wall of the insert (due to the surface tension), creating a space or a thinner barrier for the cells to pass. Have you tested different volumes of Matrigel in the insert? I use 30 ul for a Corning insert.
Good luck,
Priscila
Hi Priscila,
The inserts I'm using already come with the matrigel. I don't know if the problem is because only the cells on the edge invade the matrigel (those are the ones that I see after staining with crystal violet in the bottom) or because when I seed the cells they all go to the edge of the insert (and there's no cells in the middle from the beginning).
Hi Sofia, I'm familiar with this issue.
I had a problem with uniformal cell migration when the meniscus in the upper chamber was close to the surface. The surface tension pushes the cells to the periphery.
The issue was solved by adding more media to the upper chamber so the meniscus does not come near the upper chamber surface.
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