My question is about using Manual Tracking plugin in imageJ but before explaining my problem please let me know if you know any other method to measure cell migration using ImageJ. So, here is my problem:
I need to track cells to quantify their migration in x and y direction. I used “Manual Tracking” plugin which is quit cool but I need help in how to use it. When opening the plugin, first under Parameters section you have to define your Time Interval and x/y calibration. Can anyone explain me what are these two parameters. I ran my experiment for 96 hours using time-lapse microscope for every 20 minutes. Also my cells dimensions are nearly 20 Micrometer. Considering these numbers, what should I put for “Time Interval” and “x/y calibration”?
Also is there any way to separately calculate for x direction and y direction velocities? As I see in the outputs, Manual Tracking only gives the velocity between two frames but how we can calculates velocity components in x and y directions?