Turku Bioscience Zeiss LSM780 User Manual Download | Manualshive

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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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Summary of Contents for 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 ...

Page 2: ... Start the ZEN software 6 6 Using the fluorescence microscope 7 6 1 Visualizing the sample in ocular mode 7 6 2 Using the confocal microscope settings 8 6 2 1 Reusing settings from previous experiments 8 6 2 2 Setting up new experiment 8 6 2 3 Using Smart Setup 8 6 2 4 Manually create light paths 9 6 2 5 Visualise sample on screen 9 6 2 6 Adjust the Channels 10 6 2 7 Acquire images 10 6 3 Experime...

Page 3: ...nformation 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 The user is responsible for removing their data from the hard drive and should do so immediately Files older than 30 days are automatically wiped from the system without prior notice ...

Page 4: ...croscope environment is tidy If there are oil spills or other issues please inform CIC personnel microscopy bioscience fi 2 4 Start the heating earlier if possible When doing live cell imaging it is good to switch on the heating at least two hours in advance If someone is using the instrument ask whether it is possible to switch it on If no one is using the instrument start the instrument and swit...

Page 5: ...l touches the coverslip Next focus the sample visually through the eyepieces After imaging wipe the oil off from the objective softly with a dry lens tissue Then finish the cleaning by wiping softly with a new lens tissue moistened with isopropanol Only lens tissue may touch the objective lens When changing the sample with the immersion objective the objective must be lowered inverted microscope i...

Page 6: ...t the environment is clean If you see some issues report them to CIC personnel 4 1 1 Definite focus 4 1 2 Main switches 1 3 2 1 Main on 2 Systems PC on The computer starts 3 Components on If needed plug in and switch on definite Focus If definite focus remains off Zen will give an error on start up ...

Page 7: ...tems PC and Components Be sure that Argon cooling has finished you will hear fan switch off Switch off Main 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 weak...

Page 8: ...EN software Click the Start System button Choose Acquisition Switch on the lasers Start up will take a few minutes a loading bar will be visible If there are any errors contact CIC personnel Switch on the lasers you need The Diode 405 is always on ...

Page 9: ...chpad 1 Choose the Locate tab to view sample using the oculars 2 Transmitted light turn on bulb and open the shutter 3 Objective lenses 10x 0 45 dry 20x 0 80 dry 40x 1 2 Water 63x 1 2 Water 100x1 4 Oil 4 RL Reflected light shutter fluorescence 5 Fluorescence filters DAPI GFP RFP Analyser filter Polarization imaging NoneLSM Confocal imaging 1 2 3 4 5 ...

Page 10: ...ging configuration you created or open an existing configuration The Smart Setup function allows you to insert your fluorophores and the software will provide light path suggestions Choose your fluorophore from the dropdown box Fastest scans your labels with a single scan This gives the most spectral cross talk bleed through and should be avoided Best signal scans the light paths separately and is...

Page 11: ... light contains the 405nm diode 4 You must select the correct mirror to reflect the laser line toward the sample arrow 5 T PMT is used for collected transmitted light from lasers to create a wide field image The buttons mentioned here are used to visualise your sample on screen One you have selected 1 track like in the adjust channels image you can start optimizing the visualization of your sample...

Page 12: ...asing the Digital Offset brings up the background and blue pixels disappear Digital Gain increases the PMT gain but can also create more noise 1 Objective can be chosen both here and in the Locate window 2 Click Optimal for the maximum resolution for current objective and zoom level Increasing the Line Step value increases scanning speed as fewer lines are scanned Decreasing the Speed and increasi...

Page 13: ...tart Experiment button During Live scan mark the lowest and highest desired focus levels with Set First and Set Last Selecting Optimal will calculate the best interval between slices based on objective and lasers selected Alternatively interval between Z slices or number of Z slices can be manually adjusted NOTE Keeping interval spacing will alter the Z range Keeping slices will keep Z range and a...

Page 14: ...ifier bottle Place the heating insert onto the stage Place your sample into the insert Place the CO2 incubator lid onto the heating insert Make sure the air holes on the lid are facing the sample Switch on off the incubator and CO2 through the software Typically 37 C and 5 CO2 are used for normal cell imaging ...

Page 15: ...ou tell the software how often to capture and how long to wait between captures Time Series is activated in the Experiments page 11 Tick Show all to reveal more options Cycles specifies the number of times you want to run your image capture set up Interval specifies the gap of time between each captured image The combination of cycles and interval will determine how long the experiment runs ...

Page 16: ...de Check that that the HBO 100 lamp unit is on and manually open the FL shutter in the touch screen control unit 8 2 Weak signal with the Argon laser Check that the switch is in Run mode not Idle 8 3 Argon does not start The LASOS cooling unit is perhaps switched off ...

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