Parameter

Description

Use Wizard

Activated: Opens the Measured PSF Wizard when applying the function to create a PSF.

Bad Pixel Correction

Employs a fully automatic detection and removal of spurious or hot pixels (also known as stuck pixels) in an image stack which might interfere with the PSF extraction procedure. It is based on the analysis of the gray level variance in the neighborhood of each pixel in the image. It is usually recommended to leave this parameter active.

Bead Proximity

Only visible if Use Wizard is deactivated.
Defines the distance between two neighboring beads in μm. A bead is excluded from the averaging if the distance to a neighboring bead is greater than the minimum distance set. A smaller value leads to the detection off fewer beads, albeit ones that are further apart, while a larger value leads to the detection of more beads, but with the risk that beads will partially overlap. Range between 2.1 and 20 µm.

Detection Sensitivity

Only visible if Use Wizard is deactivated.
Determines the sensitivity with which beads are being detected. Range is between -5 to 5. Smaller Values lower the sensitivity excluding beads with weaker staining and lower signal to noise ratio, higher values include also beads with weaker staining.

Average Beads

Only visible if Use Wizard is deactivated.
Activated: Produces an image of an individual, averaged bead.
Deactivated: Produces an image in which each found bead is centered, but saved in the R dimension. A slider for Rotation (R dimension) is displayed on the Dimensions tab.