Zeiss Lightsheet Z.1, Operating Manual

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CHAPTER 4 - SYSTEM OPERATION
Carl Zeiss Left Tool Area and Hardware Control Tools Lightsheet Z.1
124 000000-1790-528 02/2013
3.3.14 Processing – Particle Tracking
Licensing Instrument
standard  optional LSM  ELYRA Lightsheet Z.1
Tool was exclusively developed for tracking molecules or particles having a dot-like
appearance in the image data.
The Processing Particle Tracking tool allows the
identification of single molecules and follows their
movement. Trajectories will be displayed.
In the image you have than the possibility in the
Particle Tracking tab to calculate the mean square
displacement and diffusion coefficients of the single
molecules.
Highlight the
Particle Tracking field to have access
to the
Particle Tracking
tool (Fig. 187).
Activate the image you want to process in the image
container and press the
Select
button to load the
image. Then expand the Method Parameters tool
window to have access to the
Particle Tracking
tools
(Fig. 188).
You can set different parameters to define the
molecules and the calculation of the trajectories:
−
Peak intensity to noise. Sets the signal over
noise threshold according to I-M > SS x PI (I =
intensity of particle; M = mean intensity of
image frame; SD = standard deviation of mage
frame; PI = peak intensity to noise).
−
Cut off. Sets a filter to suppress molecules that
deviate from the ideal Gauss shape, for example
it there is a background structure.
−
Feature mask size [pixel]. Determines the
mask used to cut out the particle.
−
Link range [frames]. Sets the lower limit in
frames over which the particle has to be
detected.
−
Displacement [pixel]. Sets the upper limit of
distance the particle is lowed to have from one
frame to the next.
−
Trajectory Length [pixel]. Sets the lower limit
in the trajectory length a particle must have
over the observed time period in order to be
considered a particle.
Fig. 187 Processing – Particle Tracking tool
selected
Fig. 188 Processing – Particle Tracking tool
field