Track object and numbers

Hi,

we successfully create a pipeline to identify our cells and track them (see attached pipeline). The biological goal is to measure the intensity of the green channel over the time in the single cell(following the same cell even if it divide). The red marker will help us to identify the nuclei. As we are not using a very high content system and we want to be sure to identify single cell and the cells are often tightly close to each other, we introduce a Manual Edited Object where we can modify the objects. After the modification we choose to assign to the object new numbers in order to identify from the excel file which cells is where.

the problem is: when we split manually one cell that the software couldn’t identify correctly, sometimes it reassign a different number sometimes it keeps the same number in the tracked object…(see attached file example 1)When I reached the end of my time points I have to redefine many objects manually but then they are all tracked as the same number and I cannot know who is who in the excel file.(see attached file splitobject t9)

we are not really interested in “tracking” the cells as anyway we have to check them manually at the end anyway but (as far as we know) only with the tracking we can visualize the number of the object on the picture so we can figure out who is who on the excel. If this is correct how can we have different numbers for each object in the tracked cells?

I also attached a sequence of 2 timepoint for green and red channel (but we used single tiff then)

thank you very much for your help

Valeria








XY3.cpproj (837 KB)

(Sorry for the delay in response…)

I think part of the problem you are seeing is that you are using TrackObjects for something that it’s not suited for: assignment of a unique object label. However, what TrackObjects does in the case of (what it sees as) an object split, is that it assigns the same label as the ancestor to the progeny. Since you say that you are interested in tracking the cells, this is not the behavior that you want.

I think the module you really want is DisplayDataOnImage, under the “Data Tools” category. Select EditedObjects as the input,“Number” as the category, and “ObjectNumber” as the measurement, to display the unique object label on the image of choice. This overlay image can be saved with SaveImages for downstream use.

Hope this helps!
-Mark