ArticleNature photonics2025
Multispectral live-cell imaging with uncompromised spatiotemporal resolution.
Article in Nature photonics, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- SPIFFI enables single-shot super-resolution and multidimensional imaging.Nature methods · 2026Article
- Wavelength-Division Holographic Multiplexing via Cascaded Metasurfaces and THz-Spaced Microresonator Frequency Combs.Advanced materials (Deerfield Beach, Fla.) · 2026Article
- Deep-Learning-Driven High-Fidelity In Vivo Hyperspectral Fluorescence Imaging Under Extreme Photon-Limited Conditions.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2026Article
- High-fidelity super-resolution microscopy datasets spanning multispectral to hyperspectral domains via diffractive optics.Scientific data · 2026Article
- Simulating Multicolor Super-Resolution Imaging Using an RGB Camera.Computational and structural biotechnology journal · 2026Article
- Mechanisms of COPII coat assembly and cargo recognition in the secretory pathway.Nature reviews. Molecular cell biology · 2025Review
- Drug Design and Delivery for Intracellular Bacteria: Emerging Paradigms.Drug development research · 2025Review
- Dual-Channel Event Microscopy for Ultrafast Biological Imaging.bioRxiv : the preprint server for biology · 2025Article
- Article
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Authors and funding
13 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Multispectral imaging is an established method to extend the number of colours usable in fluorescence imaging beyond the typical limit of three or four. However, standard approaches are poorly suited to live-cell imaging owing to the need to separate light into many spectral channels, and unmixing algorithms struggle with low signal-to-noise ratio data. Here we introduce an approach for multispectral imaging in live cells that comprises an iterative spectral unmixing algorithm and eight-channel camera-based image-acquisition hardware. This enables the accurate unmixing of low signal-to-noise ratio datasets captured at video rates, while maintaining diffraction-limited spatial resolution. We use this approach on a commercial spinning-disk confocal microscope and a home-built oblique-plane light-sheet microscope to image one to seven spectrally distinct fluorophore species simultaneously, using both fluorescent protein fusions and small-molecule dyes. We further develop protein-binding proteins (minibinders), labelled with organic fluorophores, and use these in combination with our multispectral imaging approach to study the endosomal trafficking of cell-surface receptors at endogenous levels.
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Registered trials
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