Zeiss LSM 880 Operating Manual page 318

ZEISS
Multichannel Unmixing:
When this option is chosen. the unmixing algorithm is applied to a multi-channel image (up to 10
channels) without the use of reference spectra.
The calculation of residuals and the subtraction of background based on a background spectrum are not
available. For heavily overlapping signals this method will yield poor results.
Avoid saturation of fluorescence signal in the data set to be unmixed. Saturation will generate a
high signal in the residual channel. To get the best unmixing results, define an extra background
channel, if possible.
Reference Spectra used for Linear unmixing of the Lambda stacks processed in the Unmixing View
are stored with the resulting image and can be re-loaded using the Processing → Linear
unmixing tool with the Read parameters from image button. This facilitates processing of a series
of images or Lambda stacks all with the same parameters / reference spectra.
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CHAPTER 1 - SYSTEM OPERATION
Left Tool Area and Hardware Control Tools
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LSM 880
10/2014 V_01