Is it possible to use the Relate objects module on after the enhance or suppress module? When I used the relate objects module after enhance or suppress it will only let me select the original objects and not the enhanced versions (which derives and incorrect count).
EnhanceOrSuppressFeatures is an image processing module, it doesn’t actually work on objects; you should use IdentifyPrimaryObjects on the processed image to create new objects. Does that help?
So my current pipeline is as follows:
Enhance or suppress features (on the first channel) > Identify primary objects > Enhance or suppress features (on second channel) > identify primary objects > Measure correlation > relate objects > export to spreadsheet
Do you mean I need to make an additional identify primary objects module for the enhanced images so that the redefined objects can be recognized?
Thanks so much!
Sorry, I’m having trouble following what your goal is here. Can you give some more background, example images, your pipeline etc? The pipeline you posted seems very reasonable (enhance the image and then identify objects in channel one, enhance the image and then and identify objects in channel two, then relate them), so I’m not sure exactly what isn’t happening as you expected without some more info. Thanks!
I’m doing a validation in which I compare lysotracker red dye to LAMP-GFP engineered HeLa cells. I’m doing a colocalization analysis but also need to know the total number of overlapping objects so that I can compare them to each object set respectively.
If you’re looking to count the colocalized spots from each channel, I think rather than RelateObjects you actually want the MaskObjects module. Otherwise, can you explain in more detail what you want to do? Again, example images and your pipeline to this point will help with that.