Zeiss LSM 880, Operating Manual

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CHAPTER 1 - SYSTEM OPERATION
ZEISS Left Tool Area and Hardware Control Tools LSM 880
366 000000-2071-464 10/2014 V_01
Adjust Strength checkbox:
If you have selected the Nearest Neighbor algorithm or Fast Iterative, the set strength manually checkbox
is always activated.
With the other Algorithm options, you can activate or deactivate this option using the check box next to
Set strength manually . When activated, you can enter the degree of restoration using the slider.
To achieve strong contrast enhancement, move the slider towards Strong.
To achieve less contrast enhancement, move the slider towards Weak.
If the setting is too strong, image noise may be intensified and other artifacts, such as "ringing", may
appear.
If
Set strength manually is deactivated, it determines the restoration strength for optimum image
quality automatically. The restoration strength is inversely proportional to the strength of the so-called
regularization. This is determined automatically with the help of Generalized Cross Validation (GCV).
To expand the options for
Corrections
, use the black arrow-head button. Here you can find three
additional corrections which can be activated or deactivated using the adjacent check boxes.
Lamp Flicker checkbox
: Analyzes the total brightness of each z-plane. In the event of non-constant
deviations in the total brightness between neighboring planes, a compensation factor is taken into
account. Activate this function if you have acquired your images using an old fluorescent lamp that
exhibits strong fluctuations in brightness.
Decay : Corrects changes in brightness (e.g. bleaching) of the sample during acquisition of the Z-Stack.
This function should only be activated for widefield images. Use it if your sample undergoes strong
bleaching during acquisition.
Background
Correction checkbox: Analyzes the background component in the image and removes it
before the DCV calculation. This can prevent background noise being intensified during DCV.
On the PSF settings tab (Fig. 520), you can see and
change all key parameters for generating a theoretically
calculated Point Spread Function ("PSF").
If an image is used that is not registered and not
fused, a point spread function (PSF) is used that is
specific for single view images (single view PSF) for
the Lightsheet system. If an image is loaded that
has already been registered and fused, a specific
PSF is used for this dataset. This multiview PSF is
calculated using the information on how many
views and which rotations were used to acquire
this particular dataset, taken from the image
metadata.
Ordinarily, images that have been acquired using ZEN
(stored as *.czi) automatically contain all system
parameters and image information, meaning that you do
not have to configure any settings on this page. Most
parameters are therefore grayed-out in the display. It is
possible, however, that as a result of an incorrect
microscope configuration, values may not be present or
may be incorrect. You can change these here. The
correction of spherical aberration can also be set here.
Fig. 520 Processing – Lightsheet
Processing. Multiview
Processing, Deconvolution,
PSF settings