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The laser doesn't automatically come on when you press Acquire, which is why you need to turn the laser on in the Light Control area. Multiple Acquisition Setup handles this betterautomatically switches the laser on when acquisition starts, which is one reason why I prefer it.
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By default NimOS performs molecular localisation on data as it is acquired. If you don't want it to do that, tick the box that says Disable real-time localization. |
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- 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, adjust the 635 laser power to 100% and make sure the button above the percentage slider is clicked to activate the laser (text is should be white, colour is should be bright). Set the exposure and number of images.
- Ignore the Priority and AutoFocus and Z Reference tabs. They are not important for conventional 2D dSTORM.
In the Saving tab, specify the Dataset Tag and the Acquisition Tag (see Acquisition Control above for an explanation of the tags). If you are logged into your User login all files will be automatically saved in your folder.
Note By default NimOS performs molecular localisation on data as it is acquired. If you don't want it to do that, tick the box that says Disable real-time localization.
Click the Start button at the bottom to begin the acquisition.
Note Once Multiple Acquisition Setup has run the lasers in Light Control will be greyed-out. You need to click the button that says 'Enable active lasers' before imaging again.
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