We are having some questions related to the accuracy with which TrackMate does the sub-pixel particle localization.
"To improve the localization accuracy, and extra step is taken to yield a sub-pixel localization of the spots. The position of each spot is recalculated using a simple parabolic interpolation scheme, as in [Lowe]"
We could not find the [Lowe] reference on the wiki; is it maybe missing? In addition, maybe it could be good to add a few words about what the “parabolic interpolation scheme” is?
“The RMSE on particle position is the worst here over the 4 scenarios. This is the obvious consequence of the particle shape, which is asymmetric and elongated, when the LoG detector expects bright blobs. Still, this does not affect tracking results.”
We were in fact wondering if the LoG and the DoG filter are taking into account the different width of the PSF in x&y and z; e.g., by using anisotropic Gaussian filters. Here, it reads like as if it does not. Could you please comment on this?
The background of our questions is that we would like to use TrackMate for very precise measurements of the distances of gene loci from fluorescence in situ hybridization staining; the signals here are diffraction limited spots, which basically have the shape of the PSF; typical distances are sub-micrometer.
Thank you very much,