I am currently trying to consistently track cells in a zvi movie file that contains approximately 350 images. This is for a migration assay to determine how far cells move toward a stimulus, as well as how linear their trajectory is. Ultimately, I’d like to be able to calculate the velocity of as many cells as possible in this movie file. The cell cytoplasm has been stained, which I realize can, unfortunately, complicate things, as well as the fact that my cells are of a high density. I have been using TrackObjects, as well as other modules to improve the image. I have gotten it to work decently, but I would still like to see an improvement in how well the objects are tracked. The linearity of an object can only be accurately tracked (I think) if the identification is consistent. My problem is that the identification of the cells doesn’t stay consistent for the entire 350 images for many of the cells. In one image, the cell may be labeled as 8, and then in the next, it might get merged with another object or may simply disappear. I was wondering if someone might be able to provide some advice on how to improve my pipeline, if possible.
Crop (I remove the back portion of the cells. I’m mainly interested in the first few rows of cells)
EnhanceOrSuppress features (To improve edges)
Apply Threshold (To remove background)
Identify Primary Objects
Export to Spreadsheet
Conserve memory (to hopefully make everything go a little faster)
Any help at all would be appreciated. I have attached the actual pipeline. I can email a small sample sequence if necessary but I can’t upload it here since the forum won’t accept zvi file uploads.
Thanks for any and all help,
Time Lapse Velocity Pipeline.cp (9.07 KB)