CHAPTER 1 - SYSTEM OPERATIONLSM 880 Left Tool Area and Hardware Control Tools ZEISS10/2014 V_01 000000-2071-464 3395.4.15 Processing – Particle TrackingLicensing Instrumentstandard optional LSM ELYRA Lightsheet Z.1Tool was exclusively developed for tracking molecules or particles having a dot-likeappearance in the image data.The Processing Particle Tracking tool allows theidentification of single molecules and follows theirmovement. Trajectories will be displayed.In the image you have than the possibility in theParticle Tracking tab to calculate the mean squaredisplacement and diffusion coefficients of the singlemolecules.Highlight the Particle Tracking field to have accessto the Particle Tracking tool (Fig. 464).Activate the image you want to process in the imagecontainer and press the Select button to load theimage. Then expand the Method Parameters toolwindow to have access to the Particle Tracking tools(Fig. 465).You can set different parameters to define themolecules and the calculation of the trajectories:− Peak intensity to noise. Sets the signal overnoise threshold according to I-M > SS x PI (I =intensity of particle; M = mean intensity ofimage frame; SD = standard deviation of mageframe; PI = peak intensity to noise).− Cut off. Sets a filter to suppress molecules thatdeviate from the ideal Gauss shape, for exampleit there is a background structure.− Feature mask size [pixel]. Determines themask used to cut out the particle.− Link range [frames]. Sets the lower limit inframes over which the particle has to bedetected.− Displacement [pixel]. Sets the upper limit ofdistance the particle is lowed to have from oneframe to the next.− Trajectory Length [pixel]. Sets the lower limitin the trajectory length a particle must haveover the observed time period in order to beconsidered a particle.Fig. 464 Processing – Particle Trackingtool selectedFig. 465 Processing – Particle Trackingtool field
