Zeiss LSM 700 Quick Manual
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Zeiss LSM 700 Quick Manual

Laser scanning microscope
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M i c r o s c o p y f r o m C a r l Z e i s s

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  • Page 1 M i c r o s c o p y f r o m C a r l Z e i s s...

  • Page 3: Table Of Contents

    Switching off the system ..................... 25 Introduction This LSM 700 Quick Guide describes the basic operation of the LSM 700 Laser Scanning Microscope with the ZEN 2012 software. The purpose of this document is to guide the user to get started with the system as quick as possible in order to obtain some first images from his samples.

  • Page 4: Starting The System

    Starting the System Switching on the LSM 700 system Turn on the LSM 700 system via the switch- operated multipoint connectors (power strips 1 and 2) on the system. This switches on all system components except attached scanning stages, the microscope and its illumination (HBO 100, XBO 75 or HXP 120).
  • Page 5 (Fig. 3). Fig. 3 ZEN Main Application window at Startup (a) and the LSM 700 Startup window (b and c) In the small startup window, choose either to start the system online (Start System hardware for acquiring new images) or in Image Processing mode to edit already existing images. Toggle the little...
  • 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 01/2013...

  • 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 2012 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 in case 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 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 2012 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, click the Show all tools tick box and open the Laser tool.

  • Page 12: Setting Up The Microscope

    Setting up the microscope Changing between direct observation and laser scanning mode On the Locate tab, the basic switch between manual operation of the microscope (observation of the specimen with the eye) and laser scanning mode is located. Click on the Locate tab to access the microscope controls. Pressing the Oculars Online button sets the Ocular tool active –...
  • Page 13 Setting up the microscope and storing settings Click on the Locate tab for direct observation; press the Oculars Online button for your actions to take effect immediately. Then open the Ocular tool to configure the components of your microscope like filters, shutters or objectives (Fig.
  • Page 14 Setting the microscope for reflected light Click on the Reflected light source icon to open the X-Cite 120 controls and turn it on. Click on the Reflected light shutter to open the shutter of the X-Cite 120 lamp / HBO100. Click on the Reflector button and select the desired filter set by clicking on it.

  • Page 15: Configuring The Beam Path And Lasers

    Configuring the beam path and lasers 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. Click on the Smart Setup button to open the smart setup window.
  • Page 16 Fig. 15 Proposals panel of the Smart Setup tool 01/2013...
  • Page 17 Motifs The feature Motifs is part of the Smart Setup panel. Like in photography, common imaging conditions with typical requirements also exist in laser scanning microscopy. The different Motifs are presets of scanning parameters (pixels size, pinhole diameter, scan speeds etc.) addressing such conditions (Fig.
  • Page 18 Open the Light Path tool in the Setup Manager tool group and the Channels tool in the Acquisition Parameter tool group to setup the imaging configuration. De-activate the Show all mode. The open Light Path is shown in Fig. 17. Fig. 17 Light Path tool for a single track (LSM 700) 01/2013...
  • Page 19 Settings for track configuration in Channel mode Select Channel mode if necessary (Fig. 17). Click on the LSM button in the light path tool (Fig. 17). The Light Path tool displays the selected track configuration which is used for the scan procedure. You can change the settings of the light path tool using the following functional elements: Activation / deactivation of the excitation wavelengths (check box) and setting of excitation intensities (slider).
  • Page 20 Settings for multiple track configurations in Channel Mode Multiple track set-ups for sequential scanning can be defined as one configuration (Channel Mode Configuration), to be stored under any name, reloaded or deleted. The maximum of four tracks with up to three channels can be defined simultaneously and then scanned one after the other.

  • Page 21: Scanning An Image

    Scanning an image Setting the parameters for scanning Select the Acquisition Mode tool from the Left Tool Area (Fig. 20). 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 22 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. For averaging, select the Line or Frame mode in the Acquisition Mode tool. Select the number of lines or frames to average.
  • Page 23 Image acquisition Once you have set up your parameter as defined in the above section, you can acquire a frame image of your specimen. Set Exposure, Live, Continuous or Snap buttons to start the scanning procedure to acquire an image. Scanned images shown...
  • Page 24 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 25 Scanning a Z-Stack Select Z-Stack in the main tools area. Open the Z-Stack tool in the Left Tool Area. Click on the button in the Action Button area. A continuous XY-scan of the set focus position will be performed. Use the focus drive of the microscope to focus on the upper position of the specimen area where the Z-Stack is to start.

  • Page 26: Storing And Exporting Image Data

    The WINDOWS Save As window (Fig. 28) appears. 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 27: Switching Off The System

    If using the Axio Observer microscope stand, push the power button at the left hand side to switch off the microscope. Turn off the LSM 700 system via the switch-operated two multipoint connectors on the system. Switch off the HXP 120 / HBO 100 or XBO 75 lamps and, if necessary, the scanning stage and other attached components.

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