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- Click the View button in the Acquisition Control area to begin a live feed from the acquisition camera.
- There will be two live images, which correspond to two simultaneous colour channels. For dSTORM using a far-red dye you only need Channel 1, so you can untick Channel 0 in the Image Display Options on the left. For dyes emitting wavelengths lower than far-red you will need Channel 0 to be ticked (but possibly not Channel 1 if you are not using a far red dye).
- The live camera image will only show noise until you illuminate the sample with a laser. Change the percentage power in the Light Control area by either clicking on the spinner or typing in a value and pressing the Return key. The laser power in the image below is set to 100% for dSTORM acquisition, but it is best to use a lower power like 10% to avoid photobleaching while setting up your specimen. Click on the button above the laser (e.g. in this case the red 640 button) to switch on the illumination.
- Focus on your specimen. If you are using Z lock you may need to Z Lock is NOT activated then you should change the value in the Z: field to move the focus. If Z lock is active the Z: position cannot be changed, so you should adjust the Z offset as explained in the section on focusing. Both the Z field and Z offset are highlighted with a red outline in the Position Control image below.
- Once you're in focus turn off the laser so you don’t photobleach the specimen.
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