Abstract
Early detection of retinal molecular biomarkers is crucial for addressing the unmet clinical need to prevent irreversible neural tissue damage in ophthalmic and neurodegenerative diseases. Among emerging molecular sensing techniques, non-resonant Raman spectroscopy stands out as a naturally label-free and noninvasive method, offering rich biochemical information. However, in vivo detection of non-resonant Raman spectra from retinal tissue has proven to be challenging so far. Previous studies have reported conflicting results, likely due to overwhelming pigment autofluorescence. In this study, we identified the optic nerve head as the optimal retinal location for acquiring non-resonant Raman spectra in the molecular fingerprint region. Through longitudinal intra-subject measurements, we revealed dynamic changes in the molecular composition. Furthermore, a comparative study across age groups enabled the identification of molecular alterations associated with aging. These findings establish a critical foundation for utilizing non-resonant Raman spectroscopy as an early diagnostic tool for the detection of molecular biomarkers associated with ophthalmic and neurodegenerative diseases.
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Data availability
The source data underlying graphs can be obtained from Supplementary Data 1. All other data are available from the corresponding author (or other sources, as applicable) on reasonable request.
Code availability
The raw data and analysis code to generate the results in this study is available at https://doi.org/10.6084/m9.figshare.3123027475.
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Acknowledgements
The authors acknowledge: A. Amelink and P. Bussink from TNO, P. de Bettignies from HORIBA France SAS for technical support; C. Stiebing from Leibniz-IPHT and A. Krause from the Medical University of Vienna for discussions on Raman signal processing; H. Sattmann, A. Hodul and B. Rosenauer from the Medical University of Vienna for electrical and mechanical support; S. Rentz-Chorherr, M. Martin, T. Schneider and K. Memarpour from the Medical University of Vienna for support related to the medical device approval; M.C. Nguyen for the illustration in Fig. 1A. This work was carried out in the framework of project MOON. The project has received funding from: European Union’s Horizon 2020 research and innovation program under grant agreement No 732969 (It is an initiative of the Photonics Public Private Partnership. www.photonics21.org), CD-Labor OPTRAMED (CD10260501), Carl Zeiss Labor (UE60506007) and Medical Scientific Fund of the Mayor of the City of Vienna.
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R.A.L., W.D., M.Kem., M.Sch. and J.P. conceptualized the project. R.S. designed, built and integrated the OCT/RS module. M.Sal. developed the OCT acquisition software. M.E. and W.Dj. conceptualized and assisted the integration between the IR fundus imaging module and the OCT/RS module. V.S. conceptualized and assisted the integration of the RS module. R.S., M.Ken. and R.A.L. carried out the investigation and experimental work. R.S. performed the data analysis, visualization and interpretation. R.A.L., M.A. and A.U. assisted the data analysis. H.S. and A.P. recruited the subjects and provided clinical support. R.A.L., T.S. and W.D. supervised the study. R.S. wrote the original manuscript. All authors reviewed and edited the manuscript.
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M.E. and M.Kem. are employed by Carl Zeiss Meditec AG; V.S. was employed by HORIBA France SAS; T.S. is employed by Carl Zeiss Meditec, Inc; All other authors declare that they have no competing interests.
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Sentosa, R., Kendrisic, M., Salas, M. et al. Label-free in vivo molecular profiling of the human retina by non-resonant Raman spectroscopy. Commun Biol (2026). https://doi.org/10.1038/s42003-026-09744-2
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DOI: https://doi.org/10.1038/s42003-026-09744-2


