Nikon SD (Irchel) 1: Start Up

Start the computer below the table.

Start the microscope controller below the optical table

Start the spinning disk unit

Start the both cameras

Follow this guide if you need to turn on environmental control for live cell imaging.

Localize the water dispenser

Check if there is enough water in the bottle.

If not, find a bottle on the wall and fill the water dispenser.

If you see that there is almost no water left in the bottle, please let the ZMB staff know!

Pres Ctrl + Alt + Del and log in with your CORE credentials

Star the NIS-Elements SW

Star the software either in single or dual camera mode.

For dual camera mode you can follow this guide

Open the black environmental control box to access the stage.

Turn on the light in the box, if needed.

Make sure that the objective is in the lowest position. If you are not sure, engage the escape button and reset the focus position by holding that for another 2-3 sec.

Move the condensor arm to the back.

Choose the frame insert from the tower - drawer Okolab frames

Place the insert into the stage and ensure that the red dot on the insert aligns with the red dot on the stage in the same corner.

Mount your sample and move the stage that the objective is below the sample.

Choose the 4x Overview OC.

Go Live

Press the Autoscale button (Histogram with Play icon.) This will ensure that the Histogram will be scaled and you won't miss the sample even with a weak signal.

Alternatively, you can also select 4x objective and WF fluorescence channel for the overview if BF does not provide good contrast for sample localisation

Go up with the objective rotating the knob towards yourself.

The number of arrows indicates you the speed of movement. (3 arrows = fastest)

You can change the speed with this button

Start with 3 or 2 arrows and go up carefully, until you will hear gentle beeping with which microscope indicates you are near to the focal plane. Then change the speed to 1 arrow and find your focus.

Once you focused on the slide/sample, localize the Sample Navigation tab

Choose from the sample holder - if you cannot find the exact same one, choose from the most similar one.

Check different plane for different holders

Choose Current settings and Current Z position - this will use the last used OC, which you used for focusing in previous step

Click on Preview

Microscope will acquire a tilescan, which you can use later for the navigation in the sample