CHAPTER 1 - SYSTEM OPERATIONLSM 880 Left Tool Area and Hardware Control Tools ZEISS10/2014 V_01 000000-2071-464 3415.4.16 Processing – Experimental PSFLicensing Instrument standard optional LSM ELYRA Lightsheet Z.1Tool accepts Z-Stacks. See full description of tool for details.This function allows determining the point-spreadfunction (PSF) of sub-resolution beads and usingthis experimentally determined PSF as input forStructured Illumination processing.Select the Experimental PSF tab to select theExperimental PSF Processing tool (Fig. 467).Activate the image (a 3D stack of sub resolutionbeads) you want to process in the image containerand press the Select button for Input Image toload the image. Then expand the MethodParameters tool window to have access to settingparameters (Fig. 468).You can load a 3D stack of a widefieldimage or a non-processed raw data SIMimage. LSM image formats are notaccepted to determine the ExperimentalPSF.You have the possibility to automaticaly computethe Experimental PSF by choosing Automatic fromMode drop down menu (Fig. 469).If you want to process manually and set qualityparameters, select Manual (Fig. 470).Pressing the Default button will load Standardvalues. Data will only be reused, if the result imageis selected. Than pressing the Reuse button willload the values of the selected image. The optionsSymmetrical and Damp Edge are not included inthe Reuse function.When the Symmetrical check box is ticked, aradial symmetrization of the PSF will be performed.That means that a perfectly round PSF iscalculated, even out of an oval PSF.When the Damp edge check box is ticked, theedge of the PSF will be smoothed (with a smoothtransition to zero at the edge).Fig. 467 Processing – Experimental PSF toolFig. 468 Experimental PSF – automatic modeFig. 469 Mode drop down menu
