Automatic Component Extraction - Zeiss LSM 880 Operating Manual

LSM 880 Center Screen Area / Image Containers - Display and Image Analysis
6.17.1

Automatic Component Extraction

The use of this function permits the automatic
search for the individual reference spectra in a
Lambda stack.
• Load or acquire a Lambda stack data set.
• Click on the Unmixing View tab.
• Click on the
Unmixing View Option control tab (Fig. 692).
• Select the number of components in the sample
and click Ok (Fig. 691). If a subset of existing
spectra has been already defined by manual
ROIs or by spectra from the spectra database,
the number of remaining components is
selected for ACE.
• Review the results in graph.
• When obtained spectra appear reasonable,
unmix the data
button (Fig. 692).
A new and spectrally unmixed multichannel
−
image document is created (as many
channels as previously defined components).
Benefit and limitations of the ACE method:
Little to moderate spatial overlap of emission signals is required to obtain good results.
Because it relies on reference spectra extracted from the multi-channel / Lambda stack image itself, ACE
could unmix on wrong assumptions and produce poor data.
However, it provides a solution for Emission Fingerprinting in those cases where reference spectra are not
accessible via single-labeled controls.
If the resulting data is carefully checked for plausibility and quantification of the results is avoided carried
out with the appropriate caution, this often is a helpful tool when working with spectrally overlapping
dyes.
10/2014 V_01
CHAPTER 1 - SYSTEM OPERATION
button in the
set by clicking the
000000-2071-464
Fig. 691 ACE, number
selection window
Fig. 692 Automatic Component Extraction
function
ZEISS
of components
499