1
Recording of a lambda stack
The illustration shows an 8-channel
image of a cell culture transfected
with GFP and YFP. Each image shows
the mean wavelength of the channel.
2
Definition of reference spectra
The reference spectra were obtained by
means of lambda stacks of cells single-
marked with GFP and YFP, respectively.
Top: Lambda-coded projections of a cell
marked with GFP (left) or YFP (right).
Bottom: Reference spectra for GFP
(green) and YFP (red).
3
Linear Unmixing
Using the reference spectra from the
lambda stack, the
Linear Unmixing
function generates a two-channel image,
in which each channel represents only
one of the two fluorochromes.
Top: GFP
Center: YFP
Bottom: Both channels superimposed
17
The 3 Steps of Emission Fingerprinting
GFP
YFP
Intensity
4000
3000
2000
1000
0
500
510 520 530 540 550 560 570
Emission wavelength in nm
