Microscopy with ultraviolet surface excitation for rapid slide-free histology

Abstract

Histological examination of tissues is central to the diagnosis and management of neoplasms and many other diseases and is a foundational technique for preclinical and basic research. However, commonly used bright-field microscopy requires prior preparation of micrometre-thick tissue sections mounted on glass slides—a process that can require hours or days, contributes to cost and delays access to critical information. Here, we introduce a simple, non-destructive slide-free technique that, within minutes, provides high-resolution diagnostic histological images resembling those obtained from conventional haematoxylin and eosin histology. The approach, which we named microscopy with ultraviolet surface excitation (MUSE), can also generate shape and colour-contrast information. MUSE relies on ~280 nm ultraviolet light to restrict the excitation of conventional fluorescent stains to tissue surfaces and it has no significant effects on downstream molecular assays (including fluorescence in situ hybridization and RNA sequencing). MUSE promises to improve the speed and efficiency of patient care in both state-of-the-art and low-resource settings and to provide opportunities for rapid histology in research.

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Acknowledgements

We acknowledge L. Brandi, A. Datta-Mitra, T. McBroom, A. Adelaja, A. Krueger and L. Martinez for helping with the sample preparation and imaging, J. Wilson and D. Peabody for assisting with the tissue procurement, and E. Hillman for providing critical feedback. This work was partially supported by UC Davis Department of Pathology and Laboratory Medicine start-up funds, a UC Davis Science Translation and Innovative Research grant and an unrestricted gift from Agilent Technologies. This work was performed in part under the auspices of the US Department of Energy by the Lawrence Livermore National Laboratory under contract DE-AC52-07NA27344.

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R.L. and S.G.D. developed the original observations of tissue imaging using fluorescing stains under UV LED excitation. F.F., S.G.D. and R.L. designed and fabricated the MUSE microscope based on the original design by S.G.D. and conducted the wavelength-dependent depth measurements. Z.T.H. was responsible for the colour-mapping software and maintaining the github repository. R.L., F.F., M.T., A.T. and J.A.K. performed the experiments on sample preparation, staining and comparisons with traditional histology. R.L., M.T., J.A.K. and M.L. designed and performed the comparison of MUSE and traditional histology in the central nervous system cases. R.L. and A.T. designed and performed the comparison of MUSE tissues with traditional histology, which were viewed and interpreted by J.B. and A.D.B. R.L., F.F., M.L. and J.D.M. designed and performed the FISH and RNA-Seq experiments. All authors discussed and interpreted the results and wrote and edited the paper.

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Correspondence to Richard Levenson.

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Competing interests

R.L. and S.G.D. are co-founders of a start-up company, Muse Microscopy, which is involved in commercializing MUSE technology. The other authors declare no competing financial interests.

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Fereidouni, F., Harmany, Z.T., Tian, M. et al. Microscopy with ultraviolet surface excitation for rapid slide-free histology. Nat Biomed Eng 1, 957–966 (2017). https://doi.org/10.1038/s41551-017-0165-y

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