Any changes performed with this tab are immediately updated in the FRAP view (diagram and table).

Parameter

Description

Toolbar

Offers tools to draw regions of interest into the image. These regions are then used for analysis.

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Select

Activates the selection mode that enables you to select the graphic elements in the image area.

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Clone

Activates the cloning mode that you can use to create an identical copy of the last graphic element drawn in by simply clicking anywhere into the image area. To exit this mode, either switch back to the selection mode or press the ESC key.

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Draw Rectangle

Enables you to draw a rectangular region of interest (ROI) into the image.

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Draw Circle

Enables you to draw a circle.

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Draw Ellipse

Enables you to draw a elliptical region.

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Draw Spline Contour

Enables you to draw a spline contour. You can either define the corner points by a series of clicks or you can trace a contour by keeping the left mouse key pressed. Close this contour by right-clicking. Corners are always rounded with this tool.

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Draw Polygon

Enables you to draw a polygonal region.

Parameter

Description

Fit Formula

Selects the mathematical model (mono or double exponential model) for data fitting.

Diagram Line

Selects the line thickness of the fit curve(S) in the diagram.

Normalize

Activated: Displays normalized values.
Deactivated: Displays the absolute values.

The table of the intensity values and the fit diagram are updated accordingly.

Table

Activated: Displays an additional table which shows the intensity values (for each channel) per time point. The values are corrected based on the background and reference region, if applied.

Diagram Marker

Selects the time point markers of the fit curve(s) in the fit diagram.

Time Unit

Selects the time units of the fit diagram, the fit data table and the intensity table.

Fit Range

Defines the data range for the fit algorithm.

Photofading Factor

The photofading factor is either calculated from the current reference region(s) or loaded from a previous experiment. The input field shows the currently applied value.

The photofading factor = reference(t)/reference(t=0) and then fitted by: intensity(t) = exp(-kappa * t).

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Use

Activated: Uses the photofading factor for the data fitting algorithm. In the input field, you can enter the photofading factor.

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Load

Opens a file browser to load the photofading factor from an .xml file.

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Save

Opens a file browser to save the current photofading factor as an .xml file.

Region Table

Displays the regions drawn into the image.

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#

Displays the ID of the region.

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Type

Displays the type of the region.

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Group 1, 2, 3

Allows you to select more than one region for analysis and to group them. The mean intensity values for the combined regions are used as data input.

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Background

Activated: Defines the region of interest which represents the mean background intensity that should be used for data correction. The mean intensity value of the background region is subtracted from the data prior to fitting.

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Ref.

Activated: Defines the region of interest which represents the fluorescence intensity of a reference area that has not been bleached and has not been affected by the bleach event. The mean intensity within this region is used to correct the data at each time point for any bleaching artifact that occurred during the imaging process. For this the data is divided by [reference(t)/reference(t=0)].