• Transmitted light
• Fluorescence
• Live-cell imaging
• Incubator (Temperature & CO₂)

{+}>70% \[410-800\]+

{+}>80% \[500-800\]+

{+}>90% \[450-750\]+

BF: Bright-field
DIC: Interference contrast

• Turn on the computer (#1)
• If required, turn on the incubation power bar
• Turn on the camera and laser power bar on the left of the microscope (#2)
• Turn on the microscope power bar on the right of the microscope (#3)
• Use your UdeM credentials to log in to Windows
  

• Start the Zen Blue software

1. If open, close the Zen Blue software and wait for it to close completely (up to 30 seconds)
2. On the Desktop open the Documentation folder
3. Double-click Settings for Axio-Observer Z1
4. A script will run and a black window will appear briefly
5. You can then reopen the Zen Blue software

1. Save your data
2. Close the software Zen Blue
3. Transfer your data to the D: drive (Data Storage) or to your external drive and delete it from the local C: drive
4. If used, turn of the incubation power bar
5. If used, clean oil lenses with lens cleaner and paper
6. Turn off the microscope power bar on the right of the microscope (#3)
7. Turn off the camera and laser power bar on the left of the microscope (#2)
8. Turn off the computer

• 488nm laser power loss:  alignement

(FIXED) Problem with the FRAP module:
At first could not modulate the laser intensity (yet could not find the spot on the field), rebooted and then the FRAP fiber does not carry any light (even 488)
==> Laser manipulation module was too far in x and y (noticeable as a tilt and a gap between both parts of the module).

Corrected Trigger Module configuration for the BackCam

• reassigned each camera with its own trigger module in MTB config 

• DualCam calibration parameters set, using beads
• Realigned Lasers:
  • Laser output measured in spinning disk mode at the sample with 10x objective, 1x Extended tubelens, 100% laser power and ALL lasers ON:
    • 405nm: 0.70 mW before => 0.66 mW after
      • with laser 488 OFF: 0.81 mW before => 0.83 mW after
    • 488nm: 3.75 mW before => 4.17 mW after  
      • with laser 405 OFF: 3.95 mW before => 4.33 mW after
    • 561nm: 2.1 mW before => 2.37 mW after 
    • 639nm: 0.9 mW before => 1.1 mW after 
• NOTE: AOTF2 for FRAP

AOTF2 calibration
For FRAP, the illumination is strongest with the AOTF2 at 60% and not 100%
·         Error message when switching between FRAP & SD illumination
AOTF2 Error message
FRAP/SD illumination switching triggers an error message " 'MTBAOTF2LaserLine6' is not supported by the current hardware"

• Zen 2.6 updated hotfix 12
• Microsoft Windows update
• Laser output measured in spinning disk mode at the sample with 10x objective, 1x Extended tubelens, 100% laser power and ALL lasers ON:
  • 405nm: 1.08 mW StdDev 0.01mW

o    488nm: 3.3 mW  StdDev <0.01mW (3.7 mW if laser 405 is OFF)
405nm and 488nm Interactions
When both 405nm and 488nm laser are ON simultaneously, the output power is decreased by 11%.

• • 561nm: 2.6 mW StdDev <0.01mW
  • 639nm: 1.37 mW StDev 0.02 mW

·         Control for vibrations during construction work on the 4th floor

• Error when the definite focus is used (unexpected error the definite focus did not respond)
• To solve it:
  • Turn off the Zen Software
  • Turn off the microscope power bar (#3)
  • Turn off the definite focus by pressing and holding the definite focus button
  • Turn on the definite focus by pressing once the definite focus button
  • Wait until the definite focus display shows "Detecting stand"
  • Turn on the microscope power bar (#3)
• Confirm that the definite focus is showing in the microscope tactile display
• Open Zen software and run an experiment using the definite focus to confirm it is fully functional

Added to wiki

• AOTF1 changed

A spinning disk is a multipoint confocal: due to the confocality, signal is restricted to the focal plane. Outside of the focal plane, the sample is not visible. It is thus easier to find the proper focal plane using the oculars (wide-field signal). Please refer to first focus" procedure on the "loading samples" tab.

Yes. This is an inverted microscope, designed to observe living samples. The spinning disk microscope is notably relied upon thanks to its lower phototoxicity and low out-of-focus background.