I'm trying to identify and track single cells within a cell cluster using the 'Cells' segmentation (Imaris 8). I'm working with 63x magnification and my cells are stained by two colors: mCherry for cytoplasm (RED) and yfp-mem for membrane (Yellow)
I managed to do so by the membrane channel following this segmentation path: Cells -> detect cells only ->detect cell boundary from cell membrane -> cell body detection...
note:here i didn't use any limitation by the other channel (the cytoplasm) - the filter type option.
Now i'm trying to do the 'Cells' segmentation by the cytoplasm channel, to check whether i get the same tracking results or not and didn't manage to get a good segmentation!
I did the following trials:
1. First i tried this segmentation path: Cells -> detect cells only ->detect cell boundary cytoplasm (using the red channel of course). Here the whole cluster was identified as on cell!
2. In addition to the previous, i used the filter type option to include the membrane channel in order to identify each and every cell in the cluster.. it didn't work
3.Tried all the above by segmenting a ROI (for an individual cell), it didn't work as well.
Would really appreciate any suggestion.
Thank you in advance,