Summary of Contents for Turku Bioscience Zeiss LSM780
- Page 1 Zeiss LSM780 User manual Cell Imaging & Cytometry CONTACT: microscopy@bioscience.fi +358 (0)20 7032561(office) 050 369 7532 (mobile) Turku Bioscience 5th floor, room 5077...
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Page 2: Table Of Contents
Contents 1. Information ..................... 1 2. Before Imaging ....................2 Check your sample with an ordinary fluorescence microscope ......2 Clean your slides ....................2 Check the environment ..................2 Start the heating earlier if possible ..............2 Cancellation ....................... 2 Unsure? ...................... -
Page 3: Information
1. Information The Cell Imaging and Cytometry Core is responsible for maintenance of this microscope. Each user must follow the CIC user policy. More information: https://bioscience.fi/services/cell-imaging Only authorized users may use the CIC instruments. Users must report any malfunction or problem to the CIC personnel. -
Page 4: Before Imaging
2. Before Imaging Check your sample with an ordinary fluorescence microscope It is a good practice to check the quality of your sample before making use of a more expensive instrument. Clean your slides Clean the remaining salt and mounting medium off the coverslip. Dirty coverslip compromises the image quality. -
Page 5: Working With Immersion Objectives
3. Working with immersion objectives The immersion medium should match the objective. The 40x and 63x objectives need water or oil with a refraction index of 1.334. For fixed samples and short time experiments, MilliQ water is appropriate ... -
Page 6: Starting Up And Shutting Down The Instrument
4. Starting up and shutting down the instrument Start up Note: Switch lights on and check that the environment is clean. If you see some issues, report them to CIC personnel. 4.1.1 Definite focus If needed, plug in and switch on definite Focus. -
Page 7: Argon Laser
4.1.3 Argon laser Turn the key on the Lasos control unit from 0 to 1. Switch the Argon stand-by switch from Idle to Run. The warm-up process will take several minutes. The green LED will lit when the laser is ready to use. Note: the Argon laser will emit light also when in Idle but the light intensity is much weaker. -
Page 8: Starting The Zen Software
5. Starting the Zen software Start the ZEN software Click the Start System button Start-up will take a few minutes; a loading bar will be visible. If there are any errors, contact CIC personnel Choose Acquisition Switch on the lasers Switch on the lasers you need. -
Page 9: Using The Fluorescence Microscope
6. Using the fluorescence microscope Visualizing the sample in ocular mode ! NOTE: the first user must manually open the RL shutter on the Zeiss touchpad! 1 Choose the Locate tab to view sample using the oculars. 2 Transmitted light turn on bulb and open the shutter. -
Page 10: Using The Confocal Microscope Settings
Using the confocal microscope settings 6.2.1 Reusing settings from previous experiments If you have earlier acquired images with desired light paths, open the image and use the Reuse button. To create new light paths, you 6.2.2 Setting up new experiment Use the Smart Setup or create them manually. -
Page 11: Manually Create Light Paths
6.2.4 Manually create light paths 1 Click + or trash button to add or delete track. 2 Ch1 is a standard photomultiplier tube (PMT). ChS1 is a very sensitive detector, for very weak/sensitive samples. Ch2 is a cooled PMT, which is more (red-) sensitive than Ch1. -
Page 12: Adjust The Channels
6.2.6 Adjust the Channels 1 Expand all shows all available tracks. 2 Laser power and activated lasers can be adjusted. Adjust pinhole to equalize the optical Section thicknesses in each channel. Do not equalize the Airy units. Gain (Master) defines gain (brightness). -
Page 13: Experiments
Experiments Experiments are for a protocol more advanced than a single z slice. They are activated by ticking desired boxes through corresponding window. Once parameters are selected they will open and new window, Z-stacks such as for z-stack. Execute experiments through Start Experiment button During Live scan, mark the lowest and... -
Page 14: Live Cell Experiments
7. Live Cell experiments Live imaging equipment set up Check that there is distilled water in the humidifier bottle. Place the heating insert onto the stage. Place your sample into the insert. Place the CO incubator lid onto the heating insert. Make sure the air holes on the lid are facing the sample. -
Page 15: Setting Up Time-Lapse
Setting up time-lapse Setting up a time-lapse mean you can allow the software to run the experiment without the need to be at the microscope. You optimized your imaging set up and then you tell the software how often to capture and how long to wait between captures. Time Series activated... -
Page 16: Troubleshooting
8. Troubleshooting Do not see fluorescence in the Locate mode Check that that the HBO 100 lamp unit is on and manually open the FL shutter in the touch-screen control unit. Weak signal with the Argon laser Check that the switch is in Run mode, not Idle. Argon does not start The LASOS cooling unit is perhaps switched off.
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