Zeiss Lightsheet Z.1, Руководство по эксплуатации

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CHAPTER 4 - SYSTEM OPERATION
Lightsheet Z.1 Left Tool Area and Hardware Control Tools Carl Zeiss
02/2013 000000-1790-528 149
Expand To Maximum Volume
: This checkbox is activated as default. In a multiview experiment not all
views might include all the information. When Expand To Maximum Volume is activated, the volume
of the resulting dataset after registration and fusion is determined and additional black pixel (x,y) and z-
slices are added to the views during registration. This prevents loss of data for later fusion.
Scaling x=y=z
: The dimensions are downsized to the smallest to have the same size. If the scaling in
Deconvolution, PSF settings (Fig. 232) is changed, the latter will override the scaling settings here.
Individual Registration For Every Time point
: If the multiview experiment has been performed over
time, this option becomes available.
−
If activated , each time point generates a transformation matrix and each time point is registered
individually. As a result, the time necessary for registration is multiplied by the number of time
points and can rise substantially. However, if the motorized movement of the sample by the system
has changed (e.g. due to thermic effects on the system: not fully warmed up at start, incubation,
air conditioning of the room), the Individual Registration For Every Time point option becomes
necessary.
−
If not activated , one transformation matrix is generated for all the following time points based on
the first time point. As a result, processing for registration has only to be done once which reduces
processing time.
Registration Options: Intensity Based Alignment
The Intensity Based Alignment option does not
need any fluorescent beads (fiducials) present in
the sample since registration is done based on
structural information within the image content. It
is therefore advisable to select the
Registration
Channel
which contains clearly defined
fluorescent structures within the image. One can
chose more than one channel for registration. For
each chosen channel and individual registration
will be performed; it is therefore advisable to use
one channel for the final registration. To test which
channel will lead to the best results, all channels
can be chosen for later comparison of the results.
Registration can be done as a two-step process
with a preceding
Pre-Registration
and afterwards
the Registration (Fig. 235).
The
Pre-Registration
performs the process in a
2D maximum intensity projection of the Z-Stacks;
while the
Registration algorithm uses the
complete 3D information of the Z-Stacks.
It is advisable to perform first a Pre-Registration to move similar structures already into close
proximity, which might facilitate the
Registration
.
Fig. 235 Processing – Lightsheet Processing.
Multiview Processing, Intensity Based
Alignment, Parameters