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- Click View in the Acquisition Control area to get 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 untick Channel 0 in the Image Display Options on the left. For dyes other than far-red you will need Channel 0.
- 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 spin buttons or typing in a value and pressing the Return key. The laser power in the image below is set to 100% 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. red 640 button) to switch on the illumination.
- Focus on your specimen. If you are using Z lock you may need to adjust the Z offset as explained in the section on focusing.
- Turn off the laser when not imaging so you don’t photobleach the specimen.
Acquiring images
- The best way start acquiring data is to use multidimensional acquisition. Click the Multiple Acquisition Setup button under the focus camera view on the left. In the Acquisition tab, click Single, set imaging mode to Normal, specify the laser power, exposure/frequency and number of images. In Saving, specify the Dataset Tag, which will be the folder where the files are stored, and the Acquisition Tag, which will be the filename. Note that the date will be automatically included in the filename.
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