Analyzing the RelateObjects Image

[jb2425 (Jackie)]

Good morning!

I am working on analyzing lysosome/endosome/vesicle phenotypes after subjecting cells to various knockdowns. Basically I only want information on the vesicles that are present within the cells that are suited for analysis. I do obtain elimination of all the vesicles in the ignored cells (such as those touching the image boundary) by using RelateObjects. However, for all subsequent measurements, I want to reference the relate objects image which isolates a single cell’s vesicles and no others (or at least assigns vesicles to cells and deletes extraneous data). It doesnt appear that there is any option to use the RelateObjects image for analysis, like area occupied, object size, etc. Instead, all measurements must be performed on the original image, in which ALL vesicles are present with no exclusions (although they are at least categorized by parent cell in excel). Can anyone help me out?

Here is my pipeline:vesicle_analysis.cppipe (13.0 KB)

also, here is an image to help clarify:

1. RelateObjects output, showing the cells isolated in the first panel, all vesicles in the second, and only the children vesicles to their assigned parents in the third. It is this third image that I want all subsequent analyses to be based on (rather than getting an output for size/shape/etc. of ALL vesicles present in the second panel.

Thanks!

Jackie

[wichaidi (Chonlarat Wichaidit)]

I’m facing a similar problem. Please let me know if you’ve found any solution. Thanks!

[Fabba123 (N0942245)]

Puzzling … I think if you place the RelateObjects module after all the measurements you want to make on your vesicles, you can at least get the “per parent mean” of vesicles size, shape, intensity… basically any vesicle measurement that precedes RelateObjects (under Cell measurements->Mean->Vesicles). It’s not the definitive answer, but maybe it will help.

[wichaidi (Chonlarat Wichaidit)]

Yes! I just realized that as well!! Thank you!