Spinning-disk W1 Zeiss troubleshooting

  •  Restart the multisocket on the left (near the laser).
  • Force it to quit by hitting Ctrl + Alt + Suppr, then “Gestionnaire des tâches”, then select “Metamorph” and click on “Forcer l’arrêt”. Wait few seconds, then restart Metamorph.
  • Check the laser pad on the left of the table : the key must be turned on “ON”, the shutter must be “Open”, and the diodes of each laseline desired must be switched on. The laser power must be above 5% on Metamorph.
    • Try to switch between channels on Metamorph to trigger the lightpath modification.
    • If the previous operations didn’t succeed, switch off Metamorph, and, at the back of the computer unplug the grey USB cable with a double red ring. Plug it back in the computer, and restart Metamorph
  • Check through the oculars that the sample is still at the right focus
  • Check that you activated the right channel, with the corresponding laser, at a decent power (>5%)
  • Click on different channels to trigger the modification of optical path for the microscope
  • Check the Z-motor defined. In the Multi-Dimensionnal Acquisition, under “Main”, next to “Use Dual Motors”, click on “Configure” to see which Z-motor is used for the Z-Stack. For the definition of the Z-stack, use the appropriate wheel depending on the chosen motor : the wheel next to the pad for the “Zeiss motor”, the wheel next to the joystick for the “ASI piezo motor”.
    •  For the Zeiss motor, if this operation doesn’t lead to a proper definition of Z-Stack, switch off Metamorph, and restart the multisocket near the computer. Wait the initialisation of the system to restart Metamorph
  • Remove and clean the slide
  • Clean the objective
  • Add immersion medium on the objective (if oil objective)
  • Put it back on focus
  • Switch off Metamorph, and the multisocket next to the computer. Wait few seconds, then restart the multisocket, and, once the initialization is done, restart Metamorph.
  • In the Multidimensionnal Acquisition, in “Main”, under “Position”, add a position and check the presence of the autofocus (“AF=yes”), on the newly created position.
    •  WARNING : the Autofocus requires a specific sample condition. It is highly recommended to validate your sample condition with member of the imaging facility prior to your autofocus experiment. 
  • Check the CO2 bottle level, to ensure it is not empty
    • Do not replace it by yourself, please contact a member of the imaging facility.
  • Check the valve at the top of the bottle, it must be open, as well as the black valve on the side of the pressure reducer, which should be open.
  • Check the pressure on the secondary pressure gauge, it should be at 1 bar. Use the metallic valve to adjust it if needed.
  • On the tactile pad, under “Microscope”, and “Incubation”, set the CO2 at 5%.
  • Check the level of water of the humidifier bottle (inside the chamber). If needed, please complete it with MilliQ water.
  •  WARNING : the Autofocus requires a specific sample condition. It is highly recommended to validate your sample condition with member of the imaging facility prior to your autofocus experiment. 
  • Wait few minutes, the system needs some time to adjust its CO2 level.
    •  If it stays at “HIGH”, try to adjust the pressure to a value lower than 1 bar
    • Please call a member of the imaging facility if the solutions above didn’t work.