Spectral cross-cumulants for multicolor super-resolved SOFI imaging

Super-resolution optical fluctuation imaging provides a resolution beyond the diffraction limit by analysing stochastic fluorescence fluctuations with higher-order statistics. Using n th order spatio-temporal cross-cumulants the spatial resolution and the sampling can be increased up to n -fold in a...

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Bibliographic Details
Published in:Nature communications 2020-06, Vol.11 (1), p.3023-3023, Article 3023
Main Authors: Grußmayer, K. S., Geissbuehler, S., Descloux, A., Lukes, T., Leutenegger, M., Radenovic, A., Lasser, T.
Format: Article
Language:English
Subjects:
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14/35
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631/1647/245/2225
639/624/1107/328/2236
639/624/1107/328/2238
639/766/930/2735
Channels
Diffraction
Domains
Eigenvalues
Fluorescence
Humanities and Social Sciences
Image resolution
Microscopes
multidisciplinary
Sampling
Science
Science (multidisciplinary)
Spatial discrimination
Spatial resolution
Spectra
Statistical analysis
Stochasticity
Vector analysis
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Summary:Super-resolution optical fluctuation imaging provides a resolution beyond the diffraction limit by analysing stochastic fluorescence fluctuations with higher-order statistics. Using n th order spatio-temporal cross-cumulants the spatial resolution and the sampling can be increased up to n -fold in all spatial dimensions. In this study, we extend the cumulant analysis into the spectral domain and propose a multicolor super-resolution scheme. The simultaneous acquisition of two spectral channels followed by spectral cross-cumulant analysis and unmixing increases the spectral sampling. The number of discriminable fluorophore species is thus not limited to the number of physical detection channels. Using two color channels, we demonstrate spectral unmixing of three fluorophore species in simulations and experiments in fixed and live cells. Based on an eigenvalue/vector analysis, we propose a scheme for an optimized spectral filter choice. Overall, our methodology provides a route for easy-to-implement multicolor sub-diffraction imaging using standard microscopes while conserving the spatial super-resolution property. Here, the authors generalize cumulant analysis by extending it into the spectral domain to allow multicolour super-resolution optical fluctuation imaging. The simultaneous acquisition of two spectral channels followed by spectral cross-cumulant analysis and unmixing allows denser spectral and spatial sampling of the super-resolved image.
ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-020-16841-1