TUNEL analysis in intestinal cell culture

I have just jump from genetics to cell biology and my first experiments are measuring the apoptosis rate in different intestinal cell cultures. I have been using TUNEL kit from Roche and I manage these kind of photos (see attached), however I don’t really know how to proceed or how to create a module to analyze the apoptosis rate and I hope someone have the module built and can help me…

Thank you very much,

Ainara



Hi Ainara,

Have you tried looking at the example pipelines located here? The worm, human cell and fruit fly cell example pipelines seem to be the closest to what you are describing.

Have a look at them to get a feel for how cellular identification and quantification works in CellProfiler. Then you can start adding/removing modules and tweaking settings in the pipeline to see what works best for you.

One note on your images: They look rather saturated, i.e, too bright in intensity, such that the nuclei edges are not recognizable. This will make cell identification problematic later. You may want to cut down on the exposure time on your microscope, or something similar.

Hope this helps!
-Mark

Thank you Mark, I will try what you proposed. I find quite difficult to make photos with good nuclei edges, I will also try to change te exposure times… Everything si too new for me!!

Thank you again,

Ainara