Zeiss LSM 880, Operating Manual

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CHAPTER 1 - SYSTEM OPERATION
ZEISS Center Screen Area / Image Containers - Display and Image Analysis LSM 880
494 000000-2071-464 10/2014 V_01
6.17 Unmixing View
In the Unmixing View , Lambda or channel data
sets can be analyzed and spectrally unmixed (for
more information see as well section
Processing –
Linear Unmixing ). In particular it allows to:
−
display the spectra corresponding to user-
defined ROIs (mean ROI intensity over
Lambda),
−
use Lambda Stacks and all types of multi-
channel images as input (only if the PMTs of
the LSM system are calibrated),
−
show the intensity values in table form and
copy table to clipboard or save as text file,
−
generate unmixed multi-channel images.
The available tools and the functionality of the four
general View Option control blocks
Dimensions
,
Display , and Graphics is the same as in the
Lambda Coded
View (see the previous section on
this view type). The two additional view-specific
Unmixing View Option
and
Unmixing Extract
View Option control blocks are shown in Fig. 686
and Fig. 687.
Any changes done with this View Option control
blocks are effective immediately.
The Image Display in the Unmixing View shows 3 panels: the intensity-over-lambda diagram with the
extraction bands (32-channel-LSM 710 and LSM 780 systems only), the table with the intensities in each
ROI over lambda, and the image display (see Fig. 688). The spectra from images which are acquired on an
LSM 700 are shown as integrated spectra. This reflects the way, how Lambda stacks are acquired with
the LSM 700 (see the respective description in the LSM 700 Manual).
To access the Unmixing analysis view type, load or acquire a Lambda stack data set or a multichannel
image and click on the
Unmixing tab.
The Unmixing View is available (

displayed) for all Lambda stacks and for all multi-channel
images acquired on hardware with calibrated detectors. This includes all LSM 710, LSM 780,
LSM 710 NLO, LSM 780 NLO, LSM 7 MP (if NDD detectors calibrated) and LSM 700 systems.
Detector calibration is a service task. This calibration allows that reference spectra can be
acquired at different detector gain settings than the multi-labeled image acquired for unmixing!
Fig. 686 Unmixing View Option control
block
Fig. 687 Unmixing - Extract - View Option
control block