Dialog window settings

Numerical Aperture

Enter the numerical aperture of the objective.

Immersion Refractive Index

Enter the refraction index of the immersion medium used. Predefined refraction indexes of different media can be selected from the pull-down menu.

Calibration

Enter the image calibration in μm/px.

Channels

Image channels produced by your camera are listed within this table. You can decide which channel(s) shall be processed by checking the check boxes next to the channel names. The emission wavelength value may be edited (except the Live De-Blur method).

Note

Brightfield channels are omitted automatically.

Use Spherical Aberration Correction

If turned on, spherical aberration of the PSF is calculated and used. Enter the Acquisition Depth (depth defining the center of the sample where the PSF is generated). Default value of the Acquisition Depth is taken from image metadata as a distance from the Z-Stack edge to the Z-Stack Home position. This distance is in most cases 1/2 of the Z-Stack depth. The only exception is the Asymmetric Z-Stack mode where the home position is explicitly defined by the experiment setting.

Adjust the number of Layers if you want to use the depth variant deconvolution. If you set Layers to a number bigger than one, the stack is internally split in the Z dimension into several sub-stacks (layers) and an aberrated PSF is computed separately for each one. For example a Z stack having a total range of 10 µm, 5 layers, and acquisition depth (set in the deconvolution dialog) of 10 µm will generate PSFs for the sub-volumes at 6 µm, 8 µm, 10 µm, 12 µm, and 14 µm.

Finally set the Sample Refractive Index value (refractive index of the medium used for sample acquisition). Default Sample Refractive Index is 1.35 (for water).

If Use Spherical Aberration Correction is turned off, PSF without any aberration is used.

Create new document

New document is created once the deconvolution process is finished.

Do not show this dialog for images with valid metadata

If this option is checked, the settings dialog is not shown when deconvolving images with valid deconvolution metadata.

A

Water/oil/air objective with a sample (red) in a medium under the coverslip having a different refractive index that the objective was designed for.

B

Water objective with the sample matching the objective design.

C

Sample with its surrounding solution having a different refractive index than the objective was designed for. In this case acquisition depth = objective working distance.