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- If necessary, boot up the laptop computer and login to the ONI account.
- Double-click the NimOS icon on the desktop to launch the acquisition and analysis software.
- Click the Acquire button in the top left and then click Connect to Microscope.
- NimOS will open the dialogue box below asking you to check the stage before it connects to the device to prevent damage.
- Push the interlock slider to the in position and lift the lid of the chamber to make sure the area around the stage is clear and that all samples have been removed. There should be two stacks of small disc magnets, one stack of three on either side of the stage. They are there to secure the specimen.
- Leave the lid open and click Confirm and Connect. There will be noise as the laser engine starts up and messages will appear in NimOS as the computer connects to the different components.
- You will see that the stage moves to very high position as part of its start-up. This is why it needs to be clear prior to initialisation.
- In the Temperature Control area on the left, set the Target Temperature to 31ºC or 37ºC and press the Enable Control button (the button says Disable Control once pressed). The Nanoimager operates at a temperature around 6 degrees warmer than ambient and the temperature control must be set slightly above that to maintain stability. This is something to take into consideration when planning applications where temperature is important.
- Once the temperature has stabilised the control can be disabled.
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- Slide the magnets off the stage. DO NOT pluck them off as the strength of the magnet could damage the stage by pulling on it too hard.
- Put a drop or two of immersion oil on the objective lens. Avoid getting bubbles in the oil as these will cause optical aberrations.
- Make sure the bottom of the sample is clean, as dirt on the coverslip will severely impede focusing later on. If the coverslip has been used before make sure you clean it well with 70% ethanol.
- Place your sample on the stage so that the area you want to image is on top of the objective lens (e.g., a coverslip mounted on a slide, or a well of a chambered cover glass).
Drop the magnets into place on either side of the specimen so that it is held down onto the stage. There are several six disc magnets, so the sample can be held down at several positions if necessary.
Note There needs to be space for the magnets on either side of the specimen, so vessels like 35 mm dishes can’t be used with the magnets.
- Once the specimen is in position, close the lid of the chamber and pull out the interlock slider, otherwise the imaging lasers won’t work.
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