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Zeiss LSM 710 Quick Manual

Laser scanning microscopes
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  • Page 1 DK/&/ &KZ /& h'EKE <ZDWW...

  • Page 3: Table Of Contents

    Switching off the system..................... 29 Introduction This LSM 710 / LSM 710 NLO and ConfoCor 3 Quick Guide describes the basic operation of the LSM 710 / LSM 710 NLO and ConfoCor 3 Laser Scanning microscopes with the ZEN 2009 software.

  • Page 4: Starting The System

    Starting the System Switching on the LSM system x Switch on the main switch (Fig. 1/1) and the safety lock (Fig. 1/2). x When set to ON the power remote switch labeled System/PC provides power to the computer. This allows use of the computer and ZEN software offline x To completely switch on the system, now press the Components switch to ON.
  • Page 5 Starting the ZEN software x Double click the ZEN 2009 icon on the WINDOWS desktop to start the Carl Zeiss LSM software. The ZEN Main Application window and the LSM 710 Startup window appear on the screen (Fig. 3) Fig. 3...
  • Page 6 Fig. 4 ZEN Main Application Window after Startup with empty image container Fig. 5 ZEN Main Application Window after Startup with several images loaded 04/2009...

  • Page 7: Introduction To Zen - Efficient Navigation

    Both types of users will appreciate the set of intuitive tools designed to make the use of a confocal microscope from Carl Zeiss easy and fast: The Show all concept ensures that tool panels are never more complex than needed. With Show all de- activated, the most commonly used tools are displayed.
  • Page 8 Fig. 7 ZEN Window Layout configuration More features of ZEN 2009 include:  The user can add more columns for tools to the Left Tool Area or detach individual tools to position them anywhere on the monitor. To add a column, drag a tool group by the title bar (e.g., Online Acquisition) to the right and a new tool column automatically opens.
  • Page 9 The ZEN software will ask you if you want to save your unsaved images when you try to close the application with unsaved images still open. There is no image database any more like in the earlier Zeiss LSM software versions. Fig. 8...
  • Page 10 Advanced data browsing is available through the ZEN File Browser (Ctrl + F or from the File Menu). The ZEN File Browser can be used like the WINDOWS program file browser. Images can be opened by double-click and image acquisition parameters are displayed with the thumbnails (Fig. 9). For more information on data browsing please refer to the detailed operating manual.
  • Page 11 Turning on the lasers ZEN 2009 operates all lasers automatically. Whenever they are used (manually or by the Smart Setup function) the lasers are turned on automatically. The Laser Life Extender function of the software shuts all lasers off if ZEN is not used for more than 15 minutes. To manually switch lasers on or off: x Click the show manual tools tickbox and open the Laser tool.

  • Page 12: Setting Up The Microscope

    Setting up the microscope Changing between direct observation and laser scanning mode The Ocular and Acquisition buttons switch between the use of the LSM and the microscope: x Click on the Ocular button to open the controls for the microscope beam path and for direct observation via the eyepieces of the binocular tube, lasers are blocked.
  • Page 13 x Use the focusing drive of the microscope to focus the object plane. x Select specimen detail by moving the stage in X and Y using the XY stage fine motion control. Setting the microscope for reflected light x Click on the Reflected Light icon to open the X-Cite 120 Controls and turn it on.

  • Page 14: Configuring The Beam Path And Lasers

    Configuring the beam path and lasers x Click the Acquisition button. Smart Setup The tool Smart Setup is an intuitive, user-friendly interface which can be used for almost all standard applications. It configures all the system hardware for a chosen set of dyes. x Click on the Smart Setup button to open the smart setup window.
  • Page 15 Fig. 15 Proposals panel of the Smart Setup tool Pressing Apply, automatically sets the hardware parameters in the displayed way for the dyes chosen. Linear Unmixing If the option is selected, the system is set in the lambda mode automatically. Pressing the Auto Exposure button will then optimize the settings of the Gain (Master) and offset for the given laser power and pinhole size.
  • Page 16 Setting up a configuration manually Simultaneous scanning of single, double and triple labeling:  Advantage: faster image acquisition  Disadvantage: potential cross-talk between channels Sequential scanning of double and triple labeling; line-by-line or frame-by-frame:  Advantage: Only one detector and one laser are switched on at any one time. This reduces cross- talk.
  • Page 17 Settings for track configuration in Channel Mode x Select Channel Mode if necessary (Fig. 17). The Light Path tool displays the selected track configuration which is used for the scan procedure. x You can change the settings of this panel using the following function elements: Fig.
  • Page 18 x For storing a new configuration click enter a desired name in the first line of the list box (Fig. 19), then click Ok to store the con- figuration. x For loading an existing configuration click then select it from the list box. x For deleting an existing configuration click then select it from the list box and confirm the Fig.

  • Page 19: Scanning An Image

    Scanning an image Setting the parameters for scanning x Select the Acquisition Mode tool from the Left Tool Area (Fig. 20). x Select the Frame Size as predefined number of pixels or enter your own values (e.g. 300 x 600) in the Acquisition Mode tool.
  • Page 20 Setting scan averaging Averaging improves the image by increasing the signal-to-noise ratio. Averaging scans can be carried out line-by-line or frame-by-frame. Frame averaging helps to reduce photo-bleaching, but does not give quite as smooth of an image. x For averaging, select the Line or Frame mode in the Acquisition Mode tool. x Select the number of lines or frames to average.
  • Page 21 Image acquisition Once you have set up your parameter as defined in the above section, you can acquire a frame image of your specimen. x Use one of the Auto Exposure, Live, Continuous or Snap buttons to start the scanning procedure to acquire an image. x Scanned images shown...
  • Page 22 The scanned image appears in a false-color presentation (Fig. 24). If the image is too bright, it appears red on the screen. Red = saturation (maximum). If the image is not bright enough, it appears blue on the screen. Blue = zero (minimum). Adjusting the laser intensity Fig.
  • Page 23 Scanning a Z stack x Select Z-Stack in the main tools area. x Open the Z Stack tool in the Left Tool Area. x Select Mode First/Last on the top of the Z-Stack tool. x Click on the button in the Action Button area.

  • Page 24: Storing And Exporting Image Data

    The WINDOWS Save As window appears. x Enter a file name and choose the appropriate image format. Note: the LSM 5 format is the native Carl Zeiss LSM image data format and contains all available extra information and hardware settings of your experiment.

  • Page 25: Using The Confocor 3 Module

    Switching off the system x Click on the File button in the Main Menu bar and then click on the Exit button to leave the ZEN 2009 software. x If any lasers are still running you should shut them off now in the pop-up window indicating the lasers still in use.

This manual is also suitable for:

Lsm 710 nloConfocor 3

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