Nikon N-STORM Manuale operativo - Pagina 37

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Calculate the parameters for calibration and then save

them as settings.

(1)

Click

[Identification Settings] to display the

dialog box and then click [Z axis Calibration].

(2)

Click [Auto Calibrate].

The calibration results (coefficients of the calibration curve

function) obtained through the procedure above are

automatically saved as internal settings in the N-STORM

software, and subsequently are automatically applied to

3D-STORM analysis.

Check the results of calibration on the graph

Clicking [Graph] in the [Z Calibration] dialog box displays

a graph showing the results of calibration. Display the

graph of [Wx/Wy vs. Z] if necessary, and check that Wx

and Wy intersect nearly at 0 on the horizontal axis and

that there is no large gap between the plot data

(distributed points) and the fitting data (lines).

Apply the results of calibration to an existing

dataset

To apply the new calibration result to an existing dataset,

apply the following procedure.

(1) Click [Save] in the [Z Calibration] dialog box to save

the calibration result as a file.

(2) Open the existing dataset.

(3) Display the [Z Calibration] dialog again and then click

[Load] to read the calibration result file to be applied.

Click [OK] twice to close the dialog box.

(4) Click

[Start STORM Analysis] to perform

analysis again.

Timing of calibration

When the objective has been switched or the

composition of the STORM buffer has been changed,

perform calibration again.

Chapter 2

STORM Microscopy Operation

15-(2)

15-(1)