Zeiss Lightsheet Z.1 Operating Manual

Contents

CHAPTER 4 - SYSTEM OPERATIONCarl Zeiss Left Tool Area and Hardware Control Tools Lightsheet Z.156 000000-1790-528 02/2013Light Sheet Illumination SettingsThe first choice to make regarding the Light Sheet illumination is given in the drop down menu anddetermines if the light sheet should only illuminate the sample from one side (Single Side) or if the lightsheet is sequentially generated from both sides (Dual Side) (configuration dependent). The third mode isDual Side when Experiment. Here a single side illumination is kept during the setup of the experiment,hence when using the Continuous or Snap action buttons, which facilitates the setup of the imagingparameters. The dual side illumination is only activated when the Start Experiment button is pressed inthe Multidimensional tool selection panel.If Single Side is chosen, the direction, Left or Right, must be marked. This part of the panel is inactivatedwhen the Dual Side illumination is selected.The Thickness of the Light Sheet at its center is displayed in μm.If Dual Side illumination is selected for each image (z-slice, time point) two frames are acquired, eachwith one illumination direction. The corresponding frames can be processed during image acquisition andcombined into one frame when activating the Online Dual Side Fusion checkbox. The raw data will notbe kept when Online Dual Side Fusion is selected, but only the resulting fused images. Or the raw datacan later be fused using the Lightsheet Processing tools in the Processing tab.Pivot Scan SettingsThe Pivot Scan (configuration dependent) is used to move the light sheet within the focal plane of thedetection optics in a wavelike motion. This results in a reduction of shadows which might otherwise becast by optically dense structures within the sample. It can be used when activating the checkbox next toPivot Scan.Acquisition area settingsInformation about the used number of Pixelswithin the frame is given as well as the dimensionsof the Image Size and the Pixel Size aredisplayed in μm. The dimensions will change whenusing the Zoom function described above. FrameSize and therefore the number of Pixels to beused can be adjusted by opening the drop downsub menu. Here you can choose either from a listof Rectangular or Square predefined cropsettings. The last five used pixel numbers fromeither list will appear in the Recent section of thesub menu. For individual adjustment you can typein the desired pixel amount for x and z next toManual Selection.When the Reset All button is pressed, the fullframe with all available pixels will be imaged. Thisbutton has no influence on the zoom settings.Fig. 77 Frame Size