Palm - Localization Precision - Zeiss LSM 880 Operating Manual

LSM 880
5.4.14.3 PALM – Localization Precision
Localization
Select
Localization Precision look up table (LUT) to an
Experimental PSF (Fig. 437).
Highlight the Image with the Experimental PSF
(should be derived from the PSFs obtained by a 4
µm stack of 100 nm beads) and press Select
(Fig. 438).
Press apply to obtain a file of the Experimental PSF
with LUT attached. The file bears the extension
"LocalizationPrecision".
Please note that the localization precision
will be estimated using a Monte-Carlo
Simulation
including noise. The generated lookup
table will then contain the localization
precision depending
parameters photon number, background
variance and Z position.
The Experimental PSF with attache LUT
should be used when fitting 3D PALM
data. If only an Experimental PSF without
LUT is used, a warning message pops up
(Fig. 439). Press No to cancel processing
or Yes if you want to continue. In this
case, only a rough estimate of the
localiszation precision will be given.
10/2014 V_01
CHAPTER 1 - SYSTEM OPERATION
Left Tool Area and Hardware Control Tools
Precision
to attach
of the experimental
on the
000000-2071-464
a
Fig. 437
PSF
three
Fig. 438
Fig. 439
Processing – PALM Localization
Precision tool selected
Processing – PALM - Localization
Precision with double-lobed
Experimental PSF loaded
Localization precision warning
ZEISS
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