Intraoperative microscopic autofluorescence detection and characterization in brain tumors using stimulated Raman histology and two-photon fluorescence

INTRODUCTION: The intrinsic autofluorescence of biological tissues interferes with the detection of fluorophores administered for fluorescence guidance, an emerging auxiliary technique in oncological surgery. Yet, autofluorescence of the human brain and its neoplasia is sparsely examined. This study...

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Autores principales: Fürtjes, Gina, Reinecke, David, von Spreckelsen, Niklas, Meißner, Anna-Katharina, Rueß, Daniel, Timmer, Marco, Freudiger, Christian, Ion-Margineanu, Adrian, Khalid, Florian, Watrinet, Konstantin, Mawrin, Christian, Chmyrov, Andriy, Goldbrunner, Roland, Bruns, Oliver, Neuschmelting, Volker
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10207900/
https://www.ncbi.nlm.nih.gov/pubmed/37234975
http://dx.doi.org/10.3389/fonc.2023.1146031
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author Fürtjes, Gina
Reinecke, David
von Spreckelsen, Niklas
Meißner, Anna-Katharina
Rueß, Daniel
Timmer, Marco
Freudiger, Christian
Ion-Margineanu, Adrian
Khalid, Florian
Watrinet, Konstantin
Mawrin, Christian
Chmyrov, Andriy
Goldbrunner, Roland
Bruns, Oliver
Neuschmelting, Volker
author_facet Fürtjes, Gina
Reinecke, David
von Spreckelsen, Niklas
Meißner, Anna-Katharina
Rueß, Daniel
Timmer, Marco
Freudiger, Christian
Ion-Margineanu, Adrian
Khalid, Florian
Watrinet, Konstantin
Mawrin, Christian
Chmyrov, Andriy
Goldbrunner, Roland
Bruns, Oliver
Neuschmelting, Volker
author_sort Fürtjes, Gina
collection PubMed
description INTRODUCTION: The intrinsic autofluorescence of biological tissues interferes with the detection of fluorophores administered for fluorescence guidance, an emerging auxiliary technique in oncological surgery. Yet, autofluorescence of the human brain and its neoplasia is sparsely examined. This study aims to assess autofluorescence of the brain and its neoplasia on a microscopic level by stimulated Raman histology (SRH) combined with two-photon fluorescence. METHODS: With this experimentally established label-free microscopy technique unprocessed tissue can be imaged and analyzed within minutes and the process is easily incorporated in the surgical workflow. In a prospective observational study, we analyzed 397 SRH and corresponding autofluorescence images of 162 samples from 81 consecutive patients that underwent brain tumor surgery. Small tissue samples were squashed on a slide for imaging. SRH and fluorescence images were acquired with a dual wavelength laser (790 nm and 1020 nm) for excitation. In these images tumor and non-tumor regions were identified by a convolutional neural network that reliably differentiates between tumor, healthy brain tissue and low quality SRH images. The identified areas were used to define regions.of- interests (ROIs) and the mean fluorescence intensity was measured. RESULTS: In healthy brain tissue, we found an increased mean autofluorescence signal in the gray (11.86, SD 2.61, n=29) compared to the white matter (5.99, SD 5.14, n=11, p<0.01) and in the cerebrum (11.83, SD 3.29, n=33) versus the cerebellum (2.82, SD 0.93, n=7, p<0.001), respectively. The signal of carcinoma metastases, meningiomas, gliomas and pituitary adenomas was significantly lower (each p<0.05) compared to the autofluorescence in the cerebrum and dura, and significantly higher (each p<0.05) compared to the cerebellum. Melanoma metastases were found to have a higher fluorescent signal (p<0.01) compared to cerebrum and cerebellum. DISCUSSION: In conclusion we found that autofluorescence in the brain varies depending on the tissue type and localization and differs significantly among various brain tumors. This needs to be considered for interpreting photon signal during fluorescence-guided brain tumor surgery.
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spelling pubmed-102079002023-05-25 Intraoperative microscopic autofluorescence detection and characterization in brain tumors using stimulated Raman histology and two-photon fluorescence Fürtjes, Gina Reinecke, David von Spreckelsen, Niklas Meißner, Anna-Katharina Rueß, Daniel Timmer, Marco Freudiger, Christian Ion-Margineanu, Adrian Khalid, Florian Watrinet, Konstantin Mawrin, Christian Chmyrov, Andriy Goldbrunner, Roland Bruns, Oliver Neuschmelting, Volker Front Oncol Oncology INTRODUCTION: The intrinsic autofluorescence of biological tissues interferes with the detection of fluorophores administered for fluorescence guidance, an emerging auxiliary technique in oncological surgery. Yet, autofluorescence of the human brain and its neoplasia is sparsely examined. This study aims to assess autofluorescence of the brain and its neoplasia on a microscopic level by stimulated Raman histology (SRH) combined with two-photon fluorescence. METHODS: With this experimentally established label-free microscopy technique unprocessed tissue can be imaged and analyzed within minutes and the process is easily incorporated in the surgical workflow. In a prospective observational study, we analyzed 397 SRH and corresponding autofluorescence images of 162 samples from 81 consecutive patients that underwent brain tumor surgery. Small tissue samples were squashed on a slide for imaging. SRH and fluorescence images were acquired with a dual wavelength laser (790 nm and 1020 nm) for excitation. In these images tumor and non-tumor regions were identified by a convolutional neural network that reliably differentiates between tumor, healthy brain tissue and low quality SRH images. The identified areas were used to define regions.of- interests (ROIs) and the mean fluorescence intensity was measured. RESULTS: In healthy brain tissue, we found an increased mean autofluorescence signal in the gray (11.86, SD 2.61, n=29) compared to the white matter (5.99, SD 5.14, n=11, p<0.01) and in the cerebrum (11.83, SD 3.29, n=33) versus the cerebellum (2.82, SD 0.93, n=7, p<0.001), respectively. The signal of carcinoma metastases, meningiomas, gliomas and pituitary adenomas was significantly lower (each p<0.05) compared to the autofluorescence in the cerebrum and dura, and significantly higher (each p<0.05) compared to the cerebellum. Melanoma metastases were found to have a higher fluorescent signal (p<0.01) compared to cerebrum and cerebellum. DISCUSSION: In conclusion we found that autofluorescence in the brain varies depending on the tissue type and localization and differs significantly among various brain tumors. This needs to be considered for interpreting photon signal during fluorescence-guided brain tumor surgery. Frontiers Media S.A. 2023-05-10 /pmc/articles/PMC10207900/ /pubmed/37234975 http://dx.doi.org/10.3389/fonc.2023.1146031 Text en Copyright © 2023 Fürtjes, Reinecke, von Spreckelsen, Meißner, Rueß, Timmer, Freudiger, Ion-Margineanu, Khalid, Watrinet, Mawrin, Chmyrov, Goldbrunner, Bruns and Neuschmelting https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Oncology
Fürtjes, Gina
Reinecke, David
von Spreckelsen, Niklas
Meißner, Anna-Katharina
Rueß, Daniel
Timmer, Marco
Freudiger, Christian
Ion-Margineanu, Adrian
Khalid, Florian
Watrinet, Konstantin
Mawrin, Christian
Chmyrov, Andriy
Goldbrunner, Roland
Bruns, Oliver
Neuschmelting, Volker
Intraoperative microscopic autofluorescence detection and characterization in brain tumors using stimulated Raman histology and two-photon fluorescence
title Intraoperative microscopic autofluorescence detection and characterization in brain tumors using stimulated Raman histology and two-photon fluorescence
title_full Intraoperative microscopic autofluorescence detection and characterization in brain tumors using stimulated Raman histology and two-photon fluorescence
title_fullStr Intraoperative microscopic autofluorescence detection and characterization in brain tumors using stimulated Raman histology and two-photon fluorescence
title_full_unstemmed Intraoperative microscopic autofluorescence detection and characterization in brain tumors using stimulated Raman histology and two-photon fluorescence
title_short Intraoperative microscopic autofluorescence detection and characterization in brain tumors using stimulated Raman histology and two-photon fluorescence
title_sort intraoperative microscopic autofluorescence detection and characterization in brain tumors using stimulated raman histology and two-photon fluorescence
topic Oncology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10207900/
https://www.ncbi.nlm.nih.gov/pubmed/37234975
http://dx.doi.org/10.3389/fonc.2023.1146031
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