Spots detection

[Marcos Nieves]

Hi everyone!

I am working with a set of TIRF movies of bacterial cells where we want to detect and track spots, in this Dropbox link you will find two examples.

It would be great if somebody can send us a brief instructive about how to process the movies before running Spot Detector and Spot Tracking plugins. Spot Detector default settings applied to the attached examples get many false positive spots and viceversa, some spots are not detected.

I am a researcher in the Institut Pasteur of Montevideo. We remember that in the ICY course Alexadre Dufour gave us here he used freq. analysis (Fourier transform) and/or other methods to select the right threshold for spot detection, but we do not remember the sequence of steps and specific methods he used.

Could you please help us with this specific TIRF movie?

Thanks a lot

[Stephane]

Hi Marcos,

Sorry for the late answer.
I tried the spot detector on your dataset and actually i can get strong detection by using these parameters :

Image _19_44_42.tif_keep_stack.tif :
enable scale 2 with sensitivity set to 80
enable scale 3 with sensitivity set to 80

Image _19_58_04.tif :
enable scale 2 with sensitivity set to 70
enable scale 3 with sensitivity set to 70

By modifying a bit the parameters you should be able to get what you want.
As soon you have good enough detection, just be sure to check the "Export to SwimmingPool" option in the export tab of spot detector so you can see the detections result in the Spot Tracking plugin.

Best,

- Stephane