Real Time Deconvolution

Using real time deconvolution, you can preview and acquire images and Image Sets that have been improved in real time by AutoQuant deconvolution. The image is acquired following deconvolution and you do not need to acquire raw widefield images prior to deconvolution. You do not have access to the raw widefield data if you acquire your data using Real Time Deconvolution.

From the Video Preview panel, select the Settings tab then the Real Time Deconvolution settings.
From the Algorithm drop down menu, choose a deconvolution method.

The various real time deconvolution options are described below.

2D Iterative Deconvolution

This algorithm produces the highest quality results, but at the expense of processing time and frame rate.

Wiener Filter

This algorithm produces intermediate quality results, with intermediate processing time.

PSF Selection

Choose between a Derived PSF or Theoretical PSF.

With Derived PSF selected, the PSF is calculated directly from image data so there is no need to enter optical data.

With Theoretical PSF selected, accurate optical data must be entered.

Lens selection

Select Active Lens to use the objective lens (and associated calibration) currently selected in the Optical Characteristics tool group of the Capture tab for PSF calculation. Select Selected Lens to enter the calibration, numerical aperture (NA) and immersion refractive index of your imaging objective directly in the Real Time Deconvolution settings.

Note: Correctly configuring and calibrating your objectives in your Lens list in the Optical Characteristics tool group makes configuring a Theoretical PSF easy and quick.

If there is no calibration associated with your selected lens, select the correct calibration by clicking on the Calibrations button and selecting the correct calibration from the Spatial Calibration panel.

Setting λ

Select Active Source to set the emission wavelength used for Theoretical PSF calculation to the wavelength associated with the currently selected channel. Select Custom λ to directly enter a custom wavelength in nm.

Iterations

Enter the number of iterations that the deconvolution algorithm will be applied to generate the deconvolved image. More iterations will decrease your imaging frame rate.

K Factor

The K Factor setting is only required for the Wiener Filter. It defines an adjusted logarithm of the Wiener K value.