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- Make sure the microscope objective you want to use is selected (either in the software or in the microscope control unit; once you have selected an objective it will automatically rotate into position). If you are using an immersion lens make sure to add a drop of compatible immersion oil (NOTE: For a list of all available microscope objectives please see the TECHNICAL SPECIFICATIONS).
- Place a specimen in the stage inset and insert it in the microscope stage.
- Go to Locate tab to redirect the light to the ocular and locate your specimen.
- To set up the light path for transmitted light imaging turn on the white light lamp 1 (light bulb icon) and adjust its intensity to ~4V (= ~30 %) either in the software or using the black wheel at the front of the microscope stand. Open the shutter 2 and make sure that the Transmission Light filter 3 is inserted in the light path. It is also possible to narrow down the condenser aperture to improve the contrast. You should know be able to focus on your sample down the eyepiece.
- To set up the light path for epi-fluorescence imaging change the filter 3 to None (actually a quad band filter). Turn on the X-CITE LED controller by pressing ENTER AS GUEST and then TURN ON ALL LEDs and then open the shutter 4/5. The intensities of the LEDs can also be adjusted individually. You should now be able to focus on your sample using fluorescence down the eyepiece. Remember that in the epi-fluorescence mode only the centre of the field of view will be visible because the illumination path is designed for optimal laser excitation of a small area matching the camera field of view.
- Once you have your sample in focus you should ideally give it 20 – 30 min to equilibrate on the stage before you start the acquisition of your super-resolution images or otherwise undesired sample drifts may occur.