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  1. 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.
  2. Put a drop or two of immersion oil on the objective lens.
  3. Place your sample on the stage so that the area you want to image is on top of the objective lens (e.g., coverslip mounted on a slide, or well of a chambered cover glass).
  4. Drop the magnets into place on either side of the specimen so that it is held down onto the stage. There are several 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.

  5. 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.

Focusing on the specimen

  1. The Nanoimager has no eyepieces so focusing must be done using a camera image. This can be done using sample fluorescence, but I’ve found that it is a lot easier to use the focus laser, which reflects at the glass/water interface of the specimen and forms an image of a spot on the focus camera. The Focus Camera view is on the left of the NimOS GUI. You will just see camera noise unless the focus laser is switched on.
    Image Added
Info

If the focus camera view is not visible you can select ‘Show Focus Camera View’ from the Instrument menu.

  1. Click the Focus Laser button in the Light Control area on the right. You will probably see a very faint diffuse signal at the edge of the focus camera view because it is unlikely you will be in focus. You need to change the Z position in the Position Control area on the right until there is an image of a small laser spot in the camera view. I have found that I can get close to the correct focus by typing in the values

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  1. be

For an ibidi μ-slide -300

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