I tried to calculate the illuminationcorrection function and apply it on a data set of time-lapse images, but I am not sure the background was corrected.
The problem is that over time I see that the background profile is increasing, (probably due to secretion of fluorescence to the medium or other optical aberrations) but in the control FOV the background profile is stable. I also want to eliminate, by calculating this function, all the inhomogeneities in the camera or light over time.
The illumination correction function I used is:
First pipeline is to correctIlluminationCalculate using the following parameters: Background intensities, block size of 64 (images are 1024x1024 pixels and the objects are 20-80 pixels in diameter), no rescaling, All, load images and no smoothing.
For the correct illumination Apply module I chose to load the .mat image calculated in the previous pipeline and then subtract it from the original images without scaling.
After correcting the images, I got the same increasing profile in the background.
Is the way of calculating and applying the correctillumination module was right for this application?
How do you suggest eliminating this background?
Thanks in advance,