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Zeiss LSM 710 NLO Quick Manual
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Zeiss LSM 710 NLO Quick Manual

Zeiss LSM 710 NLO Quick Manual

Laser scanning microscopes
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LSM 710 / LSM 710 NLO
and ConfoCor 3
Laser Scanning Microscopes
LSM Software ZEN 2008
August 2008
M i c r o s c o p y f r o m C a r l Z e i s s
Quick Guide
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Summary of Contents for Zeiss LSM 710 NLO

  • 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 Quick Guide LSM 710 / LSM 710 NLO and ConfoCor 3 Laser Scanning Microscopes LSM Software ZEN 2008 August 2008 We make it visible.
  • Page 3 Contents Page Contents ..........................1 Introduction..........................2 Starting the System ....................... 3 Introduction to ZEN – Efficient Navigation ................6 Setting up the microscope....................11 Configuring the beam path and lasers ................13 Scanning an image....................... 16 Storing and exporting image data ..................21 Using the ConfoCor 3 module.....................
  • Page 4 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 2008 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 5 Starting the System Switching on the LSM system • Switch on the main switch (Fig. 1/1) and the safety lock (Fig. 1/2). • 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 •...
  • Page 6 Starting the ZEN software • Double click the ZEN 2008 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 ZEN Main Application window at Startup (a) and the LSM 710 Startup window (b and c) In the small startup window, choose either to start the system online ("Start System"...
  • Page 7 Fig. 4 ZEN Main Application Window after Startup with empty image container Fig. 5 ZEN Main Application Window after Startup with several images loaded 05/2008...
  • Page 8 Introduction to ZEN – Efficient Navigation ZEN - Efficient Navigation - is the new software for the LSM Systems from Carl Zeiss. With the launch of this software in 2007 Carl Zeiss sets new standards in application-friendly software for Laser Scanning Microscopy.
  • Page 9 The Pro-Basic concept ensures that tool panels are never more complex than needed. In Basic Mode, the most commonly used tools are displayed. For each tool, the user can activate Pro Mode to display and use additional functionality (Fig. 6). Fig.
  • Page 10 The ZEN software will ask you if you want to save your unsaved images when you try to close the application. Note: There is no "image database" any more like in the earlier Zeiss LSM software versions. Fig. 8 New image document in the Open Images Ares...
  • Page 11 Advanced data browsing is available through the 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 a 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 12 (Fig. 10). Remember, the argon multi-line laser has to first be put to standby for a 5 minute warm-up before it changes to on. • Zeiss recommends operating the Argon multi-line Laser at a tube current of about 6 A (∼50 % output). This is the best compromise between laser stability/power and laser life-time. (tube-current control can be found in Pro Mode).
  • Page 13 Setting up the microscope Changing between direct observation, camera detection and laser scanning mode The Ocular, Camera and LSM buttons switch the beam path and indicate which beam path is currently in use for the microscope: • Click on the Ocular button to change the microscope beam path for direct observation via the eyepieces of the binocular tube, lasers are blocked.
  • Page 14 Setting the microscope for reflected light • Click on the Reflected Light 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. •...
  • Page 15 Configuring the beam path and lasers • Click on the LSM button. Choosing a configuration Simultaneous scanning of single, double and triple labeling: − Advantage: faster image acquisition Disadvantage: Eventual cross-talk between channels − Sequential scanning of double and triple labeling; line-by-line or frame-by-frame: −...
  • Page 16 Settings for track configuration in Channel Mode • Select Channel Mode if necessary (Fig. 15). • Click on the LSM tab (Fig. 14). The Light Path tool displays the selected track configuration which is used for the scan procedure. • You can change the settings of this panel using the following function elements: Fig.
  • Page 17 • For storing a new configuration enter a desired name first line Configurations list box (Fig. 17) and click Store. • For loading an existing configuration select it from the list box and click on the Load button. • For deleting an existing configuration select it in Fig.
  • Page 18 Scanning an image Setting the parameters for scanning • Select the Acquisition Mode tool from the Left Tool Area (Fig. 18). • 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 19 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. •...
  • Page 20 Image acquisition Once you have set up your parameter as defined in the above section, you can acquire a frame image of your specimen. • Use one of the Find, Fast, Continuous, or Single buttons to start the scanning procedure to acquire an image.
  • Page 21 The scanned image appears in a false-color presentation (Fig. 22). 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 •...
  • Page 22 Scanning a Z stack • Open the Z Stack tool in the Left Tool Area. • Select Mode First/Last on the top of the Z Stack tool. • Click on the button in the Action Button area. A continuous XY-scan of the set focus position will be performed.
  • Page 23 • The WINDOWS Save As window 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 24 Using the ConfoCor 3 module • Click on the LSM button. • Use the ConfoCor 3 Tool Group in the Left Tool Area to acquire and analyze FCS data. → Fig. 28 ConfoCor 3 Tool Group Setting a configuration • Open the Measure toolbar to access experiment parameter controls.
  • Page 25 You can change the settings of this panel using the following function elements: Activation / deactivation of the excitation wavelengths (check box) and setting of excitation intensities (slider). Open the Laser Control tool via the Laser icon. Selection of the main dichroic beam splitter (HFT) or secondary dichroic beam splitter (NFT) position through selection from the relevant list box.
  • Page 26 Starting a measurement • Open the Measure toolbar to access experiment parameter controls. • Select the Acquisition controls in the Measure tool. The Light Path and Pinhole panels of the Measure window display the selected track configuration which is used for the FCS procedure and the pinhole size (see Fig. 30).
  • Page 27 Press the New button to open a new FCS diagram into an image container. If a measurement is triggered, all data are displayed in that window if highlighted. Press the Start button to trigger a measurement. All defined positions will be approached consecutively.
  • Page 28 Fig. 31 FCS Correlation diagram You have the following function elements: Activate the FCS Correlation panel to display measured data (Fig. 31). Press the View Options button to define the graph you want to display. Press the Count rate button to display the count rate trace. Press the Correlation button to display the correlation function.
  • Page 29 Press the Data Options to handle your data. Press the Save Data button to open the Save window. You can save the whole data set in an ANSI text format. Optionally you can save the raw data trace if that option was set in the FCS Options.
  • Page 30 You have the following options: Activate the FCS Fit panel to display fitted data (Fig. 32). Set the red and blue bars to define the start and end points of the curve fit window. Load a predefined model from the Model drop-down menu. You can assemble a model by pressing the Model tool in the ConfoCor tool group.
  • Page 31 Switching off the system • Click on the File button in the Main Menu bar and then click on the Exit button to leave the ZEN 2008 software. • 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 710Confocor 3