Measure objectintensity

hi I have question about the ‘meausre object inensity’ function. I’d like to use that to measure nuclear fluorescent protein expression intensities. for example, I have a group of cells with heterogeneous expression gfp and rfp in their nuclei. I stain nuclei with dapi, and got three image frames: dapi, gfp, rfp. I identify primary objects (nulcei) in the dapi frame. following that, I use measure objectintesnity module. in this module, I can choose both images and objects and get little confused by this (is this different from CP1.0). i if I choose ‘image’ as gfp and ‘object’ as nuclei, does that mean I measure the gfp signals in corresponding nuclei identified in the dapi image? if not, what does it measure? thanks.

Hi linghuang,

Yes, you seem to understand it correctly. Here’s another way of stating what MeasureObjectIntensity does. Upstream, an Identify* Module defines regions of interest (ROI, or ‘objects’), e.g. using the DAPI channel. But to measure the intensity signals within those objects, the user can choose any (or all) of the channels to measure the signals from, e.g. DAPI, GFP, etc.

Yes, this would measure the GFP intensity (actually, multiple intensity features, see the Help for the module) in each nuclei object, as defined previously using the DAPI image (assuming that’s what you used in the upstream IdentifyPrimaryObjects module).

As for CP 1.0, I can’t remember exactly (it’s been awhile!), but I believe the only difference is that now the module offers the user to choose multiple channels and objects to choose from in the same module.

Hope that helps,
David

thank you very much. that helps a lot.