Hi @pearl-ryder ,
thank you for getting back to me, I appreciate it.
This is the pipeline I used.
first attempt.cpproj (810.5 KB)
My goal is to first identify single cells from one image, such as this one; these cells were PI stained. The cells do not move during the experiment.
PI3 pos1.tif (256.4 KB)
Once the positions of the cells are marked in the PI stained image, I need the software to overlay such positions on the rest of the images, which were taken in a time-lapse manner.
The big goal is to measure fluorescence intensity over time in these single cells. As you can see from the following spreadsheet, the software identifies primary (nuclei) and secondary (cells) objects for each image from the sequence (column 2 and 3). Instead, I need the software to maintain the positions of the 376 cells identified in the first image (the PI stained image) and simply measure the increase/decrease of fluorescence on each one of the time-lapse images, on these same 376 positions. I thought I had solved this issue by adding the OverlayObjects function but it does not work.
I also use this occasion to ask you another question. Technically, I have 10 different positions for each experiment. Meaning each position contains a different PI stained image and different time-lapse images. The total number of time-lapse images is the same for each position instead and they were all taken at the same time. For example, I would have 10 images at time t=1, 10 images at t=2 etc.
My question is: is there a way to run the same analysis that I described before on this set of images all at once or do I have to run a new pipeline for each position?
I hope my explanations were clear enough.
Thank you in advance for your support, it is much appreciated!