Huygens Professional User Guide

Deconvolving my first imageand a Parameters page. You can open as many Expand viewers as you like, on the sameimage or on different images.image is not a timeseries.‘t’-slicing is notavailable.Figure 3. Two Expand Viewers opened from the same image. Left theSlicer Tab is selected, right the Projection Tab.With the Viewer’s Slicer you can scan through the slices (in different directions). Since thisexample image is not a time series, the time (t) select box is greyed.Microscopic parameters • Verify the microscopic parameters (needed for the generationof a PSF) like Numerical Aperture, Microscope type, samplesize, etc. by selecting the Parameters Tab from the Expandviewer (see Figure 3).Did you use the correct Sampling size? To find out, selectfaba32 in the Main window, by clicking on the image (thename is displayed in blue, when selected!). Now select‘Nyquist rate’ from the Edit menu and you will be informed.We see that the image is fairly good sampled in the x and y directions since the nyquist rateis near to the value of one. However, the z (the x-y slices) is undersampled (the samplingwas too course). Advised is to sample in the z-direction with a distance of 166 nm or less.Conclusion: if the Nyquist tool leaves you with values smaller then one you have undersampledyour image. Tough luck for you now, but hopefully future recordings may benefitfrom this information.(Super sampling of an image is not a problem, although your dataset becomes unnecessarilylarge without providing extra information).For now we continue with the z-undersampled demo image.The Huygens Professional User Guide 9