Calculation of mean fluoroscence intensity minus the black background



I have an image where i have seperated the nucleus from the cytoplasm. The nucleus of the cells are easy to measure because their ROIs can be defined. but while measuring the mean fluroscence of the cytoplasm the black background of the image at the back is not giving me accurate measure of the cytoplasm fluoroscence. Please suggest what can be done.


Hello Rajan_Singh,
Most of us would like to see a sample image in either PNG or tiff format, so we may better understand what is requested. Thanks,