LEFT TOOL AREA AND HARDWARE CONTROL TOOLS
LSM 710 and LSM 780
Carl Zeiss
Processing Tab
Systems
122 M60-1-0025
e
02/2010
Display channels with statistical confidence:
This option displays an additional channel per unmixed
component which shows the relative statistical error in each unmixed component channel. This statistical
uncertainty of the pixel intensity in the unmixed channel is calculated based on the (Poisson-) noise of the
acquired (input-) channels, the bandwidth and –position and the quality of the reference spectra. The
display can be interpreted best when an appropriate false color LUT is chosen. The error is displayed as
percentage of unmixed channel pixel intensity. For example: A "confidence value", i.e., pixel intensity of
50 represents a 50% statistical uncertainty of the corresponding intensity in the unmixed channel. The
higher the statistical error of the acquired pixel intensity (the shot noise), the higher is the intensity value
of this pixel.
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. See also
Automatic
Component Extraction
.
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 Lambda stacks processed in the Unmixing
View are stored with the resulting image and can be re-loaded in 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.
Fig. 129
Image Display window before unmixing in Preview
