Zeiss LSM 880, Руководство по эксплуатации

Страница: 344 / 690

CHAPTER 1 - SYSTEM OPERATION
ZEISS Left Tool Area and Hardware Control Tools LSM 880
338 000000-2071-464 10/2014 V_01
5.4.14.5 PALM – Remove PALM Outliers
The Remove PALM outliers function allows
removing all molecules that within a defined radius
will have less than the specified numbers of
neighboring molecules.
Select the
Remove PALM outliers tab to select
the
Remove PALM outliers
processing tool
(Fig. 460). 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
filter functions (Fig. 461).
You can set two parameters by using the sliders or
the spin input boxes:
Radius [nm]
. The radius defines the distance from
a localized position of a peak (molecule) in which
all peaks falling within that distance are counted.
Position threshold . If the number of counted
neighboring peaks (molecules) falls short of the set
value, the respective peak (molecule) will be
removed from the list and the PALM image.
This tool will emphasize densely labeled
structures and can help remove floating
molecules and ghost signals.
This tool works only for PALM images.
After setting the thresholds, press Apply to create
the new image. A message will be displayed telling
how many molecules have been removed from the
input PALM image (Fig. 462).
Press
Ok to have access to the newly computed
image (Fig. 463).
Please note that for 3D-PALM removing
outliers applies only in 2D for a maximum
intensity projection of the Z-Stack. Removing
outliers does not apply for the z direction.
Fig. 460 Processing – Remove PALM outliers
tool selected
Fig. 461 Processing – PALM Remove PALM
outliers
Fig. 462 Processing – PALM Remove Outliers
message box
Fig. 463 PALM images before (left) and after
(left) removal of outliers; Radius set to
3000 and Position Threshold to 100.
Only those peaks / molecules in denser
structures remain.