Living Image 3.1

8. Spectral Unmixing
8.1 Spectral Unmixing
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1. Load the image sequence.
In this example, the fluorophore is Quantum Dots 800. Images were acquired using
a 675 nm excitation filter and emission filters from 720 to 820 nm in 20 nm
increments.
2. In the Analyze tab of the Spectral Unmixing tools, put a check mark next to the emission
wavelengths that you want to include in the analysis.
Excitation
wavelength
Emission
wavelengths of
the sequence
3. In the Components drop-down list, select the
number of spectral components to unmix in the
images (the number of fluorophores +1 since
tissue autofluorescence accounts for 1
component).
For example, if the image data includes one
fluorophore, then there are two components to
unmix−the fluorophore signal and the tissue
autofluorescence.