Does anyone know of a program or plugin that can automatically quantify vesicle/organelle fusion and fission events? For example, I would like to track fluorescently-labelled lysosomes and track and quantify the number of lysosome-lysosome fusion and fission events.
Dear Roberto,
How easy this can be done will depend on the characteristics of your time series data. This includes factors such as temporal (t) and spatial (x,y,z) resolution and the clarity of your signals (signal to noise ratio; cell density).
As software I might recommend the Tracking Feature of Ilastik (find documentation here http://ilastik.org/documentation/tracking/tracking). As far as I recall it can handle splitting and merging objects. Alternatively you might try the TrackMate plug-in of imagej (https://imagej.net/TrackMate). In the latter case you can extract gap-closing, splitting and merging events more easily.
Hi Roberto,
The software I use is NIS-Elements. The HCA module does a really good job identifying most objects you can spot by eye. The tracking module will track the movement of each object (lysosomes in this case) over time as well as record how often two objects fuse into a single object or a single object splits into two.
Once you have optimized the settings, you can run this on all of your imaging files to output fission and fusion rates.
Just a heads up, Elements is not a free software package, but if you have questions about the specifics, feel free to message me.
Best,
Andy
You can talk to Warren Lee at St. Mike's (Toronto). They are doing this.
Hi, maybe late but you can try IMARIS which done easily tracking of object (but simplicity have a cost) and give stats about fusion and fission.
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