Zeiss LSM 880 Operating Manual page 343

LSM 880
Expand the Grouping field to have access to the
Grouping tool (Fig. 458).
You can set two filters to be taken into
consideration when peaks (events) are grouped:
− Off Gap. If peaks within the defined
Capture Radius will be interrupted by more
than the set value of consecutive frames,
they will not be regarded belonging to the
same molecules.
− Capture radius [pixel]. If peaks in consecutive frames are not localized within the specified radius
they are not regarded as belonging to the same molecule.
Note that the settings of the Grouping will not influence the Preview image.
Note that using Grouping in the processing tool is irreversible. Therefore, in the created PAL-
Grouping tab all functions will be greyed out and not accessible.
If all parameters and filters are set press the Apply button. This will create the PALM image in the image
container and the PALM view tab becomes available that allows further processing and analysis of the
image. The container will display PALM image (HR channel) as well as the sum widefield image (SWF
channel – not available for 3D-PALM) (Fig. 459).
Note that in contrast to the TIRF image, which is the raw image of the point spread functions
(PSFs), the sum wide-field image is the background corrected image of the PSFs and the PALM
image is a vector map of centroids.
Fig. 459
10/2014 V_01
CHAPTER 1 - SYSTEM OPERATION
Left Tool Area and Hardware Control Tools
PALM image (left) and sum widefield image (right) after
PALM Processing
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Fig. 458 Grouping panel
ZEISS
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