Filter by Intensity

Hi CP team,

I’ve been reading previous posts and also the help section (?) to understand how the filter object works. I’m interested in filtering by intensity but I am a bit confused and wondering if you can explain a bit more or have an example to illustrate?

I want to use this in my co-culture imaging assay to eliminate the false positive stained cells. (The CTRL are on either side of the plates i.e col 2 & 23 in a 384well plate) A draft of my pipeline is as below and it would be great if you can comment on it and if I should include any other steps?

-ColorToGray
-Morph
-MaskImage
-IdentifyPrimaryObjects
-ShrinkToObjectCenter
-IdentifySecondaryObjects
-MeasureObjectIntensity
-MeasureObjectSizeShape
-MeasureTexture
-MeasureGranularity
-FilterObjects
-ExporttoExpreadsheet

Do I also need to have another pipeline setup to calculate my CTRL intensity first? or can I have them together in one & how?

Many thanks,
Shu-Oi

Hi,

Filtered objects don’t retain their “parents” measurements (aka if you measure the Size/Shape of “Nuclei” and then filter to “GFPPosNuclei”, “GFPPosNuclei” will NOT have any Size/Shape measurements associated), so I would move FilterObjects up to right after MeasureObjectIntensity; if you need intensity measurements of the filtered objects, you can then add a second MeasureObjectIntensity after the filtration step. Other than that it seems reasonable!