Applying objects to a new image?


#1

For my quantitation purposes, I use three color images to define my cells:

  1. DAPI (nuclear stain, for primary objects)
  2. Texas Red maleimide stain (nonspecific stain for the whole cell, for “secondary” objects defining the whole cell)
  3. 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?


#2

Hi,

You simply need to add the “MeasureObjectIntensity” module and choose to analyze the GFP image, then choose the cytoplasm object. This will record the GFP signal in your defined cytoplasm.

Mike


#3

Thanks, I ended up figuring it out myself, but I wasn’t entirely sure that it was quantifying the GFP image and not the reference images used to make the objects.