Start-up procedures#
Start-up#
- Turn on surge protector (switch on the right).
- Turn on laser box "F XL" by rotating key clockwise.
- Turn on microscope controller (white Nikon box to right of laser box).
- Turn on CSU-X1 unit (connected to the left port of microscope) by rotating key clockwise.
-
Uncover and turn on scope body by pushing a switch on the right side of the scope at the back. (1)
- Sometimes the scope doesn't turn on properly. You'll know this is the case if it starts beeping and/or the joystick pad doesn't light up. Don't worry--just shut off the scope body (wait for the beeps to finish), and try starting again.
-
Sit at computer and start NIS-Elements by double-clicking on the NIS-Elements AR icon.
- Select your account from the drop-down menu (default: no password).
- Choose
Front Fusion
for single-camera imaging orDual Fusion
for dual-camera imaging. - Check the drawer to see the TwinCam DM's status. Ensure it's out for single-camera imaging and in for dual-camera imaging.
Video showing start-up steps 1-5
Live-cell checklist#
Allow CO2 flow and start Tokai Hit unit#
- Open the Apollo valve for CO2 by rotating counter-clockwise 90° until the yellow handle is parallel to the tubing.
Photo of CO2 line
- Turn on the Tokai Hit incubator control box by pushing the orange button.
- Check the airflow indicator and ensure the ball is at the 150 ML/MIN line.
Objective Heater (only for immersion objectives!)#
- If using an immersion objective, gently attach the objective warmer by wrapping it around the upper part of the objective.
Important
Excessive tension will damage the lens heater.
- Adjust the correction collar.
- Add a drop of Nikon Type F immersion oil to the objective.
Set-up Tokai Hit stage-top incubator#
- Ensure the Tokai Hit stage adapter is installed (do not overtighten the screws!)
- Put the Tokai Hit stage top incubator onto the stage while lifting the cables/tubing with your right hand to prevent undesirable tension on the cables/tubing.
- Add 20mL of distilled/DI water to the reservoir.
- Inspect the sample adapter for oil (and clean if needed) before inserting into the Tokai Hit stage top incubator.
- Insert your sample (or a clean dish/slide), and attach the Tokai Hit dish/slide lid. Make sure the magnetic lid is flat, or your sample will be tilted.
- If using an immersion objective, raise the objectives so that the oil and the bottom of the dish make contact.
- Put the stage top incubator lid on.
- Turn off lights, and close the door to the microscope room.
Important
- Wait >60 minutes for temperatures to stabilize before imaging.
- If using dual camera acquisition, please see Dual Camera Alignment for next steps.