For my quantitation purposes, I use three color images to define my cells:
- DAPI (nuclear stain, for primary objects)
- Texas Red maleimide stain (nonspecific stain for the whole cell, for “secondary” objects defining the whole cell)
- GFP (the channel I want to quantitate the cytoplasm of)
So far I’ve been able to use the DAPI image to detect the nuclei, then use the Texas Red image to define the “cells”, and finally use the two to get a tertiary “cytoplasm” object. What I’d like to do next is to apply the “cytoplasm” outline to the GFP image so I can quantitate object intensity there. Can anyone suggest a good way to do this?
Or, alternatively, if this functionality isn’t available–can anyone suggest a workaround?