CHAPTER 1 - SYSTEM OPERATIONLSM 880 Left Tool Area and Hardware Control Tools ZEISS10/2014 V_01 000000-2071-464 1715.2.23.2 Defining the Scan Parameters• Open the Acquisition Mode tool (Fig. 252).• Select Frame mode and a Frame Size of 512 x512 to start with.• The Optimal button automatically adjusts theframe size to match the magnification andresolution of the objective. Thus, images areacquired without collecting redundant imageinformation.• The Scan Speed is set to 9 per default. A lowerspeed produces better images as the lightcollection per pixel is higher. Use a speed from7 to 9 to begin with.• Start with the following settings on the Averaging panel:Bit depth: 8 bitsNumber: 1 (no averaging)Additional parameters like scan direction are displayed in Show all mode. Scan direction is perdefault set to unidirectional.The default settings for Acquisition Mode are suitable for good image quality and do not needto be changed for initial scanning.5.2.23.3 Defining the Detection Parameters• Activate the Channels tool (Fig. 253).• Set the Pinhole to one Airy Unit clicking 1 AU.− The pinhole diameter should be so small thatthere is still enough variation for the settingof the Gain (Master) and that sufficientimage information is still available. 1 Airy is agood value to enable a confocal fluorescenceXY-image to be obtained.− A small pinhole diameter will increase thedepth of focus, but reduce the light intensityreceived by the PMT photomultiplier (forreflection mode confocal images start with apinhole value of 0.5 Airy Units).• Use the Set Exposure button.− The system determines the settings for theGain (Master) and digital offset to match thescan speed, pinhole size and laser power.− Finally, the system builds up the image.Fig. 252 Acquisition Mode toolFig. 253 Channels tool
