Zeiss LSM 880 Operating Manual page 350

ZEISS
Once settings have been completed the Preview window displays beads that are not taken into account
as red circles and those that are used in white circles (if channel color is grey) or in their channel color (if
chanel color is other than grey) (Fig. 474). Press apply to compute the Experimental PSF. The created file
will have the extension "... .ExperimentalPSF. ...".
Fig. 474
Best results for an Experimental PSF for 3D data will be obtained, if beads are seeded sparsely,
for example 10 - 20 beads evenly distributed in a 512 x 512 image.
Fig. 475
If Apply is pressed an average PSF from all the beads taken into account will be computed (Fig. 475). For
each channel a separate PSF will be displayed. The Experimental PSF can be saved (format .czi) and
used for the Structured Illumination - Structured Illumination Processing tool in place of the
Theoretical PSF. The size and pixel size of the Experimental PSF are recalculated and the PSF is
interpolated to match the processed image.
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CHAPTER 1 - SYSTEM OPERATION
Left Tool Area and Hardware Control Tools
Preview window for bead selection using 3D PALM
Experimental PSF in Ortho view
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LSM 880
10/2014 V_01