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Hi,
I am making movies where I capture several ROI's of puncta simulatneously. These puncta disappear or diminish in intensity over time at differing rates. I'd like to use ImageJ to quantify the change in punctal intensity from the initial frame compared to the last frame (where intensity has variably diminished depending on the ROI). What would be the optimal way of doing so? Given that I'm looking at several different populations of puncta with their own rates of intensity diminishment, ideally, it would be great to graph the intensity changes as a distribution.
Another problem is that, given all of the choices in ImageJ: 1) do I deconvolve the frames and, if so, will this help the program to better identify puncta? Should this be helpful, what would be the best method of deconvolution in ImageJ? 2) what would be the optimal means of selecting thresholds to identify as many of the puncta within an ROI as possible?
Any and all help is much appreciated!
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