Hey everybody, I recently started analyzing live cells in the Operetta system. They are stained with Tubulin tracker (633) and Mito tracker (546) and no nuclei stain since I want to keep them alive after staining for some time. Now, of course I cannot use the "find nuclei" command in columbus and wanted to switch to "find cytoplasm" for normalization. But the software does not allow me to select that. I'm sure it's just a technicality, but I'm unable to find the reason. Can anyone help me out? Attached find a screenshot showcasing my issue. Thanks!!
Hi Milena Korneck,
I have never worked with that particular software, but with other image analysis softwares with similar building blocks. It could be that the cytoplasm gets detected around a first object, usually the nuclei. Or the cytoplasm gets detected by substracting a nucleus from a cell body, etc. Meaning that without having nuclei identified you might not be able to use that option. Did you try the "Find Cells" option at some point? That sounds like it could give you what you need.
If you want to inlcude a nuclear dye, have you thought about Hoechst? That can be used in live cells.
I hope I could help you somehow.
Best of luck,
Selene
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