Zeiss LSM 880, Operating Manual

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CHAPTER 1 - SYSTEM OPERATION
LSM 880 Left Tool Area and Hardware Control Tools ZEISS
10/2014 V_01 000000-2071-464 351
The
Preview
tab shows the
Input
image and a
Preview
of the output image.
By pressing
Default
the current parameter values will be stored as default values. By pressing
Reuse
the
last stored default values will be loaded.
Pressing
Attach
allows parameter values to be attached to an image. In normal case if any of the SIM
parameters will be changed, they will be applied accordingly for the processing, but may not be saved
into the image. Then if the image is saved, the values of the parameters applied for the image will be
lost. If you attach the corresponding tag to the image by pressing the
Attach
button parameter values
used to process the image will be attached to the image, even if it is not explicitly saved again. They will
then be available for further processing, in particular for Batch processing.
In the
Output
drop down menu you can set, what
kind of channels you want to display (Fig. 493). You
have the option to display
−
SR-SIM + DCV + Widefield
−
SR-SIM + DCV
−
SR-SIM + Wide Field
−
SR-SIM.
Highlight the option you want to display. Channel
designations are as follows:
SR for SR-SIM, WF for
Widefield and
D
for Deconvolution (DCV).
If the Raw Scale box is unchecked, output images
will be normalized to the maximum dynamic range.
Please note that in this case quantitative information
is lost, however, images are displayed with best
brightness and contrast. If you want to keep the
original scaling for quantitative studies, you have to
check the
Raw scale box. In this case the orifginal
dynamic range is kept.
You can select between theoretically computed or
experimentally determined PSF by choosing
Theoretical
or
Experimental
in the
PSF
drop down
menu (Fig. 494). In case Experimenta l is selected, the
PSF File button becomes available (Fig. 495). Pressing
the button will open Windows Explorer, which allows
you to select any pre-saved Experimental PSF. The file
name of the loaded file is displayed in the
PSF File
display window.
Note, if the number of the Experimental PSF
channels is less than the number of the WF or
SIM channels, the theoretical PSF will be used
for the missing channels and a warning is generated. The program also checks the match of the
excitation wavelengths between the PSF and WF / SIM channels. In case of mismatching, it
generates a warning, but the experimental PSF will still be used.
By pressing Apply the high resolution image will be computed with the current parameter values
(Fig. 496).
If a value is changed you have the possibility to press
Combine
. Then a new calculation is only done for
the changed parameter and combined with the previous calculations on the unchanged parameters,
which will save dramatically computational time. If
Apply is pressed, calculations will be done on all
parameters regardless of being changed or unchanged.
Fig. 493 Output drop down menu
Fig. 494 PSF drop down menu
Fig. 495 PSF selection menu
Fig. 496 SIM image (left) and widefield
image (right) in the Split mode
after SIM processing