Zeiss Lightsheet Z.1, Руководство по эксплуатации

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CHAPTER 4 - SYSTEM OPERATION
Carl Zeiss Left Tool Area and Hardware Control Tools Lightsheet Z.1
118 000000-1790-528 02/2013
Expand the
Localizer
field to have access to the fit
model used to localize peaks (Fig. 171). Two fit
models are available that can be selected by the
Fit
Model drop down menu (Fig. 172)
−
x,y 2D Gauss Fit
−
Mask fit
If
x,y 2D Gauss Fit is selected, the intensity
distributions of the identified peaks are fitted to a
full 2D Gaussian function. The fit in each direction
takes into account the influence of the other
direction (x,y 2D Gauss).
If
Mask Fit is selected, a Gaussian mask with a
defined PSF width is used to match as best as
possible the intensity distribution of a peak in x
and y (Fig. 173). The PSF width can be set via the
PSF Width slider or input spin box. Default value is
the theoretical calculated PSF based on the
objective lens and laser line used.
Note that with the
Mask Fit
only one predefined PSF width will be used. Computation of the Fit
will be faster for the Mask Fit , but less precise.
The settings of the
Localizer
will not influence the peak finding process and hence the preview
image.
In the Processing
PALM
function, peak finding and localization are always linked. If you want to
just view localized peaks (molcules), you might select in the PAL-Render tab of the PALM image in
the
Display
drop down menu
Centroids
.
If the
Average before Localization
check box
remains inactive, every peak (event) will be
localized independently and is regarded as being
derived from one molecule. Later on, peaks can be
grouped belonging to one molecule in the image
container in the
PAL-Grouping
tab of the image
( localization before grouping ). If the check box
is ticked, however, the system will group peaks
(events) based on the filter settings that become
available (Fig. 174) before localizing the grouped
peaks (
grouping before localization ).
When
localization before grouping
is performed, the single peaks are first localized and then the
localizations averaged. When grouping before localization is performed, than the peaks are first
averaged and then localized.
Fig. 171 Localizer Expansion field with x,y 2D
Gauss Fit selected
Fig. 172 Fit Model drop down menu
Fig. 173 Localizer Expansion field with Mask Fit
selected
Fig. 174 Localizer Expansion field with Average
before localization selected