We have a tif file that contains 73 images (sequence) that has two channels. Red and Green. We want to use the second channel with the threshold as a mask and then plot the intensity of the two channels over time.
We had managed to do it some time ago but for some reason it is not working right now. What we are doing right now is:
Select red channel -> Apply threshold
Convert to mask
Image->Stacks->Plot Z-axis profile
I’m not sure the sequence of actions is correct. Any idea on how to do this? The end result was a plot that had the intensity of each channel as a line on the same window.