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. 2023 Mar 7;13:3829. doi: 10.1038/s41598-023-30680-2

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Hyperspectral images of biopsies. (a) Color image, (b) BLUE 400, (c) Hyperspectral overlay, (d) PPIX contribution [µg/ml] per pixel. Average PPIX contribution [µg/ml] and the evaluated tissue area size are given. (1) IZ biopsy with no visible fluorescence in the microscope; hyperspectral measurement visualizes PPIX fluorescence and residual blood. (2) ST biopsy of small size with pixels of high reflectance and underestimated PPIX contribution. Microscope fluorescence rating was “none”, whereas, in hyperspectral imaging, weak fluorescence was visible (2b). (3 and 4) RABT samples of middle size, fluorescence quality “weak” in the microscope. Both showed heterogeneous PPIX distribution. (5) Tiny ST biopsy showing strong fluorescence in the surgical microscope and an artificially high PPIX contribution in hyperspectral measurement of up to 164 µg/ml.