Versatile and comprehensive hyperspectral imaging tool for molecular neuronavigation: a case study on cerebral gliomas.

Dorotea Nardini, Anam Toaha, Camilla Bonaudo, Ivan Ezhov, Angelos Artemiou, Manuel Camelia, Filippo Nozzoli, Luca Giannoni, Ilias Tachtsidis, Alessandro Della Puppa, Daniel Rueckert, Pietro Ricci, Francesco Pavone

Journal: Journal of biomedical optics 2025;30(12):126007

PMID: 41403976

Abstract

SIGNIFICANCE

Accurate and timely characterization of brain tumors remains a major challenge in neurosurgery. Current intraoperative guidance relies on preoperative imaging modalities such as magnetic resonance imaging, positron emission tomography, or computed tomography, which are essential for surgical planning but become less reliable during surgery due to brain shift. Furthermore, postoperative tumor classification depends on histopathology, which requires weeks and can delay treatment decisions. No existing tool offers real-time, label-free, and spatially resolved biomolecular information to support both intraoperative guidance and early tissue assessment.

AIM

We developed HyperProbe1.1 (HP1.1), a hyperspectral imaging system designed to acquire comprehensive molecular and metabolic information from brain tissue without the need for contrast agents or staining.

APPROACH

HP1.1 captures reflectance images across a broad range of narrow spectral bands, enabling spatial mapping of hemoglobin, cytochrome c oxidase, and oxygen saturation. In addition, ultraviolet-excited autofluorescence imaging provides information on metabolic cofactors - nicotinamide adenine dinucleotide and flavin adenine dinucleotide - relevant for tumor characterization. The system was validated using standardized phantoms and ex vivo glioma samples.

RESULTS

HP1.1 demonstrated strong performance in detecting spectral features across phantoms and in distinguishing glioma tissues of different histological grades, enabling the generation of rapid and spatially resolved molecular contrast maps.

CONCLUSIONS

By providing label-free, high-content, and rapid biomolecular imaging, HP1.1 represents a powerful platform for noninvasive tissue assessment in controlled experimental settings and paves the way for future intraoperative applications.

© 2025 The Authors.

Address: University of Florence, Department of Physics and Astronomy, Sesto Fiorentino, Italy.; European Laboratory for Non-linear Spectroscopy, Sesto Fiorentino, Italy.; University of Florence, Azienda Ospedaliero-Universitaria Careggi, Neurosurgery, Department of Neuroscience, Psychology, Pharmacology and Child Health, Florence, Italy.; Technischen Universität München, Klinikum rechts der Isar, Munich, Germany.; University College London, Department of Medical Physics and Biomedical Engineering, London, United Kingdom.; Careggi University Hospital, Histopathology and Molecular Diagnostics, Florence, Italy.; Technischen Universität München, Klinikum rechts der Isar, Munich, Germany.; Imperial College London, Department of Computing, London, United Kingdom.; Munich Center for Machine Learning (MCML), Munich, Germany.; University of Florence, Department of Physics and Astronomy, Sesto Fiorentino, Italy.; European Laboratory for Non-linear Spectroscopy, Sesto Fiorentino, Italy.; National Institute of Optics, National Research Council, Florence, Italy.
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