Nikon N-STORM Manuale operativo - Pagina 35

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Check the minimum intensity of bright points

(1)

Click

[Use Peak Statistics].

(2)

Select the darkest bright point of all those to be

identified as molecules, and move the mouse

pointer onto its center. Read and write down the

value of [Peak Height]. (This value is used as the

minimum intensity for the identification of bright

points in the next step.)

Configure the settings for the identification of bright

points.

Click

[Identification Settings].

In the dialog box, configure the settings as follows.

[Minimum Height]:

Specify the minimum intensity, checked in (2) of

the previous step.

[Maximum Height]:

[CCD Baseline]:

Select the [3D] checkbox.

Click [>>] to configure the following detailed settings.

[Minimum Width (nm)]: 200

[Maximum Width (nm)]: 700

[Initial Fit Width (nm)]:

[Max Axial Ratio]:

[Max Displacement (pix)]: 0 (This allows bright points

recorded in multiple frames to be analyzed as

separate molecules, increasing the accuracy of

calibration.)

Click [OK].

Restore the Max Displacement setting

When performing analysis except for calibration, reset

[Max Displacement (pix)] to the default value of 1.

Chapter 2

STORM Microscopy Operation

20000

100

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2.5

7-(1)

7-(2)