I’m working on some time lapse microscopy, and unfortunately my microscope suffers from drift that I can’t fix. I’m collecting z-stack images so that I have at least one image in the correct focal plane.
As my cells are moving on a bed of micropillars, it’s simple to choose the correct image as the StDev is high, as is the max. value.
Can someone point me in the right direction of how I might go about analysing a stack to select out the image with the highest StDev? Ultimately, I’d like to be able to create a 4D stack, and work through it automatically to choose the in-focus image for each time point?
Image 10 in the montage above is in focus for my application (I’m tracking the pillar movements)