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This minimizes the bleed-through between channels, mainly due to overlapping emission spectra of these fluorochromes.
We imaged 6 µm diameter double-green fluorescent microspheres (FocalCheckTM, Molecular Probes®-Invitrogen, Carlsbad CA, USA) with shell and core overlapping emission spectra (shell-green1 = 512 nm, core-green2 = 525 nm) (Figure 1.B1).
One solution is to use spectral unmixing of overlapping emission profiles.
FRET requires donor and acceptor fluorophores with overlapping emission and excitation spectra, respectively.
The thylakoid emission spectrum is highly heterogeneous and might be resolved in some partially overlapping emission bands corresponding to specific CP complexes [ 10, 28].
Commercial microarray scanners and software cannot distinguish between spectrally overlapping emission sources, and hence cannot accurately identify or correct for emissions not originating from the labeled cDNA.
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On the contrary, in the example from 18 February shown in Figure 3, there is no indication that the difference in polarization angle values might be the result of overlapping emissions, and there is also no visible gap.
The simultaneous and in situ detection of spectral information during FMT acquisition provides localized tissue specificity when calculating the relative contributions of the investigated fluorophores, thus increasing the accuracy of in vivo quantification, even with overlapping emissions.
First, they can be fluorescent components of the culture with emission spectra that overlap the emission spectrum of the analyte of interest.
It seems that the ground station detected two overlapping simultaneous emissions, each with different and almost opposite polarization angle values, thus suggesting that they exited the ionosphere in two different directions from the station (e.g., Strangeways (1986)).
Fluorescence enhancement is observed when the plasmon resonance band is excited and overlaps the emission of the fluorophore.
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