CHAPTER 1 - SYSTEM OPERATIONLSM 880 Left Tool Area and Hardware Control Tools ZEISS10/2014 V_01 000000-2071-464 3295.4.14.3 PALM – Localization PrecisionSelect Localization Precision to attach aLocalization Precision look up table (LUT) to anExperimental PSF (Fig. 437).Highlight the Image with the Experimental PSF(should be derived from the PSFs obtained by a 4μm stack of 100 nm beads) and press Select(Fig. 438).Press apply to obtain a file of the Experimental PSFwith LUT attached. The file bears the extension"LocalizationPrecision".Please note that the localization precisionwill be estimated using a Monte-CarloSimulation of the experimental PSFincluding noise. The generated lookuptable will then contain the localizationprecision depending on the threeparameters photon number, backgroundvariance and Z position.The Experimental PSF with attache LUTshould be used when fitting 3D PALMdata. If only an Experimental PSF withoutLUT is used, a warning message pops up(Fig. 439). Press No to cancel processingor Yes if you want to continue. In thiscase, only a rough estimate of thelocaliszation precision will be given.Fig. 437 Processing – PALM LocalizationPrecision tool selectedFig. 438 Processing – PALM - LocalizationPrecision with double-lobedExperimental PSF loadedFig. 439 Localization precision warning
