Zeiss LSM 880, Operating Manual

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CHAPTER 1 - SYSTEM OPERATION
LSM 880 Left Tool Area and Hardware Control Tools ZEISS
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Fast Iterative:
The "Fast Iterative" method is an iterative restoration method that uses only one iteration
per convolution step (see Meinel, E.S.: Origins of linear and nonlinear recursive restoration algorithms. J.
Opt. Soc. Am, 3 (&), 1986, 787-799). No regularization is used in this case. Due to the fast processing
and convergence after just a few iterations, this method is suitable in particular for the processing of
larger time lapse images. The results of the method can quickly lead to good results and remove most of
the out-of-focus light. They do not create quantitative brightness conditions in the image. If under-
sampled images are present, artifact formation may also result.
Constrained Iterative:
The best image quality is achieved using the iterative maximum likelihood
algorithm (see Schaefer et al.: "Generalized approach for accelerated maximum likelihood based image
restoration applied to three-dimensional fluorescence microscopy", J. of Microscopy, Vo. 204, Pt2,
November 2001, pp. 99ff.): This algorithm is able to calculate light from various focal planes back to its
place of origin. Consequently, with this method it is possible to derive the 3D structure from fluorescence
images with the correct brightness distribution and to visualize optical sections. It is also possible for
missing information to be partially restored from neighboring voxels. The spatial resolution can be
increased without artifacts up to a theoretical limit (one voxel). It is essential for Z-Stacks to have been
acquired in accordance with Nyquist. Acquiring sufficient planes above and below the structure of
interest is also imperative for achieving good results. As this is a complex mathematical method, the
calculation can take longer, depending on the image size.
If the checkbox
Enable channel selection is
activated, the settings are applied on a channel-
specific basis. This allows you to set parameters in
a targeted way for each channel. You will see a
separate colored tab for each of the channels
(Fig. 519).
In the
Normalization drop-down list, you can
specify how the data from the resulting image are
handled:
Clip: For non-registered and non-fused multiview
images, this should be used as the default setting.
It sets negative values to 0 (black). If the values
exceed the maximum possible gray value of 65636
when the calculation is performed, they are limited
to 65636 (pixel is 100% white).
Automatic:
Normalizes the output image
automatically. Not to be used for non-registered
and non-fused multiview images.
In this case, the lowest value is 0 and the highest
value is the maximum possible gray value in the
image (gray value of 65636). The maximum
available gray value range is always utilized fully in
the resulting image.
Fig. 519 Processing – Lightsheet Processing.
Multiview Processing,
Deconvolution, Enable channel
selection