ZFIN Image: Chiang et al., 2023, Fig. 1

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Figure Caption

Fig. 1

Schematic illustrating how HyU enhances analysis of multiplexed hyperspectral fluorescent signals in vivo.

a, Multicolor fluorescent biological sample (here a zebrafish embryo) is imaged in hyperspectral mode, collecting the fluorescence spectrum of each voxel in the specimen. b, HyU represents spectral data as a phasor plot, a 2D histogram of the real and imaginary Fourier components (at a single harmonic). c, Spectral denoising filters reduce the Poisson and instrumental noise on the phasor histogram, providing the first signal improvement. d–f, The phasor acts as an encoder, where each histogram bin corresponds to a number n of pixels, each with a relatively similar spectrum (e). Summing these spectra effectively averages the spectra for that phasor position. This denoising results in cleaner average spectrum for this set of pixels, which are ideally suited for analytical decomposition through unmixing algorithms (f). g, Unmixing results in images that separated into spectral components. Here, LU is used for unmixing, but HyU is compatible with any unmixing algorithm.

Acknowledgments

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