Hi everyone,
I'm using the Sartorius Incucyte SX5 for in vitro live imaging experiments and I'd like some clarification about the quantification of the fluorescent signal.
The provided software allows me to quantify some preset metrics (Total Area, Total integrated intensity, ...) and to create new analysis metrics.
I'd like to quantify the fluorescent signal and since I'm testing a cytotoxic compound (meaning I'll have less cells at the end of the treatment in the treated-wells compared to the controls - as you can see in the videos I uploaded) I want to normalize the fluorescent positivity to the area covered by the cells.
For this assay I used a cytotoxicity marker (binds the nucleic acid - green) and an Annexin V probe (NIR, Near InfraRed).
I tried creating a metric, as you can see in the attachments but I'm not sure about its validity so I'm asking you how to define the metric I need.
Thanks,
Simone
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