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1. Click on the Configuration tab.
   
2. Click the Laser Config button.
   
3. Turn on the lasers and drag the STED 592, STED 660 and STED 775 nm sliders up to 100%. Note that the White Light Laser (WLL) should automatically be at 70%. You don't need the 405 Diode on unless you will be imaging DAPI, Hoechst or other blue dyes.
   

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Activating STED and carrying out a STED alignment

1. STED is only possible using the specially optically corrected 100x HC PL APO CS2 oil lens, so you must first rotate this into position in the nose piece by selecting it in the drop-down menu in the UI.
   
2. After the 100x lens is in position you can click the Activate STED button in the top right of the left control screen. The Activate STED button will be greyed out until the 100x lens is in position.
   
3. At this stage you may be prompted to carry out an automatic STED alignment procedure. Even if you are not prompted you should carry out the STED alignment because in my experience the lasers can become misaligned over time and the alignment procedure has fixed that.

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