Hello everyone, I am trying to measure the traction forces for mouse L-cells using Traction Force Microscopy technique. I am growing cells on a 5kPa polyacrylamide gel embedded with fluorescent beads (1um), and MATLAB software for data analysis, where the traction forces are coming out to be more in unconstrained (without cells) images than the constrained (with cells) ones. This is opposite to the expected outcome. Can someone please help me in troubleshooting this problem?

Thanks