Zeiss Lightsheet Z.1 Operating Manual

Contents

CHAPTER 4 - SYSTEM OPERATIONLightsheet Z.1 Left Tool Area and Hardware Control Tools Carl Zeiss02/2013 000000-1790-528 145Adjust Strength checkbox:If you have selected the Nearest Neighbor algorithm or Fast Iterative, the set strength manually checkboxis always activated.With the other Algorithm options, you can activate or deactivate this option using the check box next toSet strength manually. When activated, you can enter the degree of restoration using the slider.To achieve strong contrast enhancement, move the slider towards Strong.To achieve less contrast enhancement, move the slider towards Weak.If the setting is too strong, image noise may be intensified and other artifacts, such as "ringing", mayappear.If Set strength manually is deactivated, it determines the restoration strength for optimum imagequality automatically. The restoration strength is inversely proportional to the strength of the so-calledregularization. This is determined automatically with the help of Generalized Cross Validation (GCV).To expand the options for Corrections, use the black arrow-head button. Here you can find threeadditional corrections which can be activated or deactivated using the adjacent check boxes.Lamp Flicker checkbox: Analyzes the total brightness of each z-plane. In the event of non-constantdeviations in the total brightness between neighboring planes, a compensation factor is taken intoaccount. Activate this function if you have acquired your images using an old fluorescent lamp thatexhibits strong fluctuations in brightness.Decay: Corrects changes in brightness (e.g. bleaching) of the sample during acquisition of the Z-Stack.This function should only be activated for widefield images. Use it if your sample undergoes strongbleaching during acquisition.Background Correction checkbox: Analyzes the background component in the image and removes itbefore the DCV calculation. This can prevent background noise being intensified during DCV.On the PSF settings tab (Fig. 232), you can see andchange all key parameters for generating a theoreticallycalculated Point Spread Function ("PSF").If an image is used that is not registered and notfused, a point spread function (PSF) is used that isspecific for single view images (single view PSF) forthe Lightsheet system. If an image is loaded thathas already been registered and fused, a specificPSF is used for this dataset. This multiview PSF iscalculated using the information on how manyviews and which rotations were used to acquirethis particular dataset, taken from the imagemetadata.Ordinarily, images that have been acquired using ZEN(stored as *.czi) automatically contain all systemparameters and image information, meaning that you donot have to configure any settings on this page. Mostparameters are therefore grayed-out in the display. It ispossible, however, that as a result of an incorrectmicroscope configuration, values may not be present ormay be incorrect. You can change these here. Thecorrection of spherical aberration can also be set here.Fig. 232 Processing – LightsheetProcessing. MultiviewProcessing, Deconvolution, PSFsettings