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2.2

Acquiring 3D-STORM (2D-STORM) Images

To perform STORM analysis, automatically repeat light stimulation and image acquisition with the appropriate laser power to

create a dataset. There are two types of mode for acquiring images: normal mode and continuous mode. In normal mode,

activation and imaging are performed in separate frames. In continuous mode, they are performed simultaneously.

The following basically describes the procedure for acquiring 3D-STORM images. Any differences from the procedure for

acquiring 2D-STORM images are noted together with the symbol "".

2.2.1

In the example below, the following two types of probe are used.

• Channel 1: Alexa405-Alexa647 dye (Alexa647 activated by Alexa405)

• Channel 2: Cy3-Alexa647 dye (Alexa647 activated by Cy3)

Calibration for 3D-STORM

Before the first 3D-STORM image is acquired, it is necessary to perform calibration. See "2.5 Calibration for

3D-STORM."

Click [Settings] in the [N-STORM] control window and

then specify the number of frames for each cycle.

[Continuous Mode]: Off

[Activation Cycle] (number of activation frames): 1

[Reporter Cycle] (number of reporter frames): 3

After making this setting, click [OK].

Select [3D-STORM] from the list at the top left of the

[N-STORM] control window.



To acquire a 2D-STORM image, select

[2D-STORM].

Chapter 2

STORM Microscopy Operation