Calculating Clathrin foci lifetimes at the plasma membrane with TrackMate

Hello community,
I have these curious thoughts in my mind about some things TrackMate can do :wink:

I was wondering if it would be possible to quantify the lifetimes of the round Clathrin foci at the plasma membrane with TrackMate from a time-lapse movie… In theory it should be possible as i have seen that TrackMate can detect these foci at the membrane. If the lifetime of these foci can be considered like a ‘track’ (as these foci slightly diffuse at the membrane), then the duration of the track should give the lifetime of the foci right? I tried to this but the tracks been detected by Trackmate are not precise as they start cross linking these foci. I was wondering if there is a way around this?

Here is a link to a Tiff stack of a Clathrin time lapse movie (2 seconds per frame) if anyone would like to have a look - https://cloud.uni-halle.de/index.php/s/yv6bfVQz1wpGtjf

Any suggestions/comments would be appreciated.
Many thanks,
Praveen

Hi @Praveen

you should be able to avoid cross-linking the tracks of neighboring foci by restricting the Linking max distance and Gap-closing max distance (e.g. in Simple LAP tracker) to a value close to zero, e.g. 0.1 pixels.

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Hi @imagejan
Thanks for your help. That seems to do the work :slightly_smiling:

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