Zeiss LSM 880, Operating Manual

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CHAPTER 1 - SYSTEM OPERATION
LSM 880 Left Tool Area and Hardware Control Tools ZEISS
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5.4.14.3 PALM – Localization Precision
Select Localization Precision to attach a
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 of the experimental PSF
including noise. The generated lookup
table will then contain the localization
precision depending on the three
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.
Fig. 437 Processing – PALM Localization
Precision tool selected
Fig. 438 Processing – PALM - Localization
Precision with double-lobed
Experimental PSF loaded
Fig. 439 Localization precision warning