18
Channel Unmixing
If the emission spectra of fluorescent markers over-
lap only slightly, the signals can be separated with
the
Channel Unmixing
function.
As raw data for unmixing in this case, it is suffi-
cient to have multichannel images in which one
of the marker dyes dominates in each channel.
Such images can be acquired without the META
Detector, i.e. with an LSM PASCAL, LSM 510 or a
CCD camera.
Channel Unmixing
also allows unmixing based on
the excitation behavior of dyes, if the raw data are
multichannel images in which the channels differ
only by their excitation wavelength.
Spectral Imaging
Double labeling of the nervous system
of a zebra fish embryo
Two-channel single-track images with emission crosstalk.
The nerve labeled with Alexa 488 can be discerned (arrows)
in the green (top) and, faintly, in the red channel (center).
Bottom: Superposition of the two channels.
Specimen: Prof. M. Bastmeyer, Friedrich Schiller University of Jena,
Germany.
The same images after Channel Unmixing.
The Alexa 488-positive nerve is visible in the green channel
(top) only but vanished from the red one (Center).
Bottom: Superposition of the two channels.
