Label-free biochemical imaging and timepoint analysis of neural organoids via deep learning-enhanced Raman microspectroscopy
File(s)
Accepted version
Author(s)
Type
Journal Article
Abstract
Three-dimensional organoids have emerged as powerful models for studying human development, disease and drug response in vitro. Yet, their analysis remains constrained by standard imaging and characterisation techniques, which are invasive, require exogenous labelling and offer limited multiplexing. Here, we present a non-invasive, label-free imaging platform that integrates Raman microspectroscopy with deep learning-based hyperspectral unmixing for unsupervised, spatially
resolved biochemical analysis of neural organoids. Our approach enables 2D and 3D mapping of cellular and subcellular structures in both cryosectioned and intact organoids, achieving improved imaging accuracy and robustness compared to conventional methods for hyperspectral analysis. Using our platform, we demonstrate volumetric imaging of a neural rosette
within a neural organoid, and interrogate changes in biochemical composition during early developmental stages in intact neural organoids, revealing spatiotemporal variations in lipids, proteins and nucleic acids. This work establishes a versatile framework for high-content, label-free (bio)chemical phenotyping with broad applications in organoid research and beyond.
resolved biochemical analysis of neural organoids. Our approach enables 2D and 3D mapping of cellular and subcellular structures in both cryosectioned and intact organoids, achieving improved imaging accuracy and robustness compared to conventional methods for hyperspectral analysis. Using our platform, we demonstrate volumetric imaging of a neural rosette
within a neural organoid, and interrogate changes in biochemical composition during early developmental stages in intact neural organoids, revealing spatiotemporal variations in lipids, proteins and nucleic acids. This work establishes a versatile framework for high-content, label-free (bio)chemical phenotyping with broad applications in organoid research and beyond.
Date Acceptance
2026-06-17
Citation
Science Advances
ISSN
2375-2548
Publisher
American Association for the Advancement of Science (AAAS)
Journal / Book Title
Science Advances
Copyright Statement
Copyright This paper is embargoed until publication. Once published the Version of Record (VoR) will be available on immediate open access.
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Publication Status
Accepted