Nikon N-STORM - Startup and basic set-up

Getting the system ready

Startup procedure

1. Turn on the Laser Power Strip (1).

• Turn on the switch that says "TURN ON!".
• Turn on the switch(es) corresponding to the laser box(es) that you will need. For example, if you only need the TIRF lasers, only turn on the "TIRF" switch.

2. Turn on the appropriate laser box(es) by turning the key to the "On" position.

• Turn on ONLY the laser boxes that you intend to use. For example, if you only need the TIRF lasers, only turn on the TIRF laser box.
• If using the FRAP/LUN4 lasers, press the grey buttons on the laser box for ONLY the laser lines you intend to use. Please, leave the others in their standby (grey) state. The TIRF laser box does not have buttons; just the key.

3.    Turn on the Microscope Power Strip (2). This also turns on the Nikon N-STORM arm and the cameras.

4. Turn the dial to your desired camera system on the Camera Box (3).

• Ensure that the dial is either pointing toward the top (A) or the bottom (D) position, NOT (B) or (C).

5. Check the light path for your desired camera system.

• For the Andor(s), remove the TwinCam filter, if you do not need it.
• For the Hamamatsu, remove the DoubleHelix phase mask, if you do not need it.

6. Start Nikon Elements.

• Select the appropriate Nikon Elements software based on your desired camera setup and capabilities.

7. Make beautiful images and don't forget to add them to the CU Up Close submissions folder on the Desktop. :)

Environmental chamber start-up procedure

1. If it is not already on, turn on the Environmental Power Strip (4). 

2. Turn on the Oko Controller (5).

• Set the controller to your desired temperature and CO2 %.

3. Open the CO2 tank valve (top knob only), located near the door (6).

4. Check the DI water in the humidifier, located to the left of the microscope. 

• You should be able to see the DI water just above the black heating unit (this model will not bubble).
• If it is low, gently unscrew the red cap and add more DI water (located in the “lobby” area of the core facility next to the sink) until it reaches the top of the black heating unit. Excess water can damage the humidity sensor, so please be mindful!

5. Attach the humidified gas line to the environmental chamber stage insert at the bronze inlet.

6. If using the 100X TIRF objective, adjust the temperature correction collar to match the temperature that you will be using.

7. Close all doors to the microscope enclosure.

8. Allow 30 – 60 minutes to reach a stable temperature. 

Inserting the sample stage inserts

Depending on your experiment you may choose a different plate. Stage inserts are found on the shelf near the CO₂ tanks.

Insert the stage insert into the stage by resting one of the short ends of the stage insert on the flange/ledge on the inside of the stage. Carefully, lower the other edge of the insert to rest on the other flange/ledge of the stage.

You can turn on the light in the environmental chamber using the pedal switch on the floor under the microscope. The intensity can be adjusted with the white knob on top of the environmental chamber. If there are any oil stains on the equipment, please clean up with ethanol before starting your experiment.

N-STORM microscope features

This microscope offers several capabilities: it has 3 light sources and 3 cameras. This manual will describe how to use the TIRF lasers but you could also use the FRAP lasers or the brightfield mode with the Lumencor SOLA Light Engine.

The Nikon motorized TIRF arm controls the TIRF angle.

There is a sCMOS camera, Hamamatsu Quest 2 with 2304x2304 pixels, and two EMCCD cameras, Andor iXon 897 with 512x512 pixels. You can use a single EMCCD camera or both of them for simultaneous two-channel imaging.