Cargando…

Optical Characterization of Neurosurgical Operating Microscopes: Quantitative Fluorescence and Assessment of PpIX Photobleaching

Protoporphyrin IX (PpIX) induced by 5-aminolevulinic acid (5-ALA) is increasingly used as a fluorescent marker for fluorescence-guided resection of malignant gliomas. Understanding how the properties of the excitation light source and PpIX fluorescence interact with the surgical microscope is critic...

Descripción completa

Detalles Bibliográficos
Autores principales: Belykh, Evgenii, Miller, Eric J., Patel, Arpan A., Bozkurt, Baran, Yağmurlu, Kaan, Robinson, Timothy R., Nakaji, Peter, Spetzler, Robert F., Lawton, Michael T., Nelson, Leonard Y., Seibel, Eric J., Preul, Mark C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6105612/
https://www.ncbi.nlm.nih.gov/pubmed/30135440
http://dx.doi.org/10.1038/s41598-018-30247-6
_version_ 1783349659710783488
author Belykh, Evgenii
Miller, Eric J.
Patel, Arpan A.
Bozkurt, Baran
Yağmurlu, Kaan
Robinson, Timothy R.
Nakaji, Peter
Spetzler, Robert F.
Lawton, Michael T.
Nelson, Leonard Y.
Seibel, Eric J.
Preul, Mark C.
author_facet Belykh, Evgenii
Miller, Eric J.
Patel, Arpan A.
Bozkurt, Baran
Yağmurlu, Kaan
Robinson, Timothy R.
Nakaji, Peter
Spetzler, Robert F.
Lawton, Michael T.
Nelson, Leonard Y.
Seibel, Eric J.
Preul, Mark C.
author_sort Belykh, Evgenii
collection PubMed
description Protoporphyrin IX (PpIX) induced by 5-aminolevulinic acid (5-ALA) is increasingly used as a fluorescent marker for fluorescence-guided resection of malignant gliomas. Understanding how the properties of the excitation light source and PpIX fluorescence interact with the surgical microscope is critical for effective use of the fluorescence-guided tumor resection technique. In this study, we performed a detailed assessment of the intensity of the emitted blue light and white light and the light beam profile of clinical grade operating microscopes used for PpIX visualization. These measurements revealed both recognized fluorescence photobleaching limitations and unrecognized limitations that may alter quantitative observations of PpIX fluorescence obtained with the operating microscope with potential impact on research and clinical uses. We also evaluated the optical properties of a photostable fluorescent standard with an excitation-emission profile similar to PpIX. In addition, we measured the time-dependent dynamics of 5-ALA-induced PpIX fluorescence in an animal glioma model. Finally, we developed a ratiometric method for quantification of the PpIX fluorescence that uses the photostable fluorescent standard to normalize PpIX fluorescence intensity. This method increases accuracy and allows reproducible and direct comparability of the measurements from multiple samples.
format Online
Article
Text
id pubmed-6105612
institution National Center for Biotechnology Information
language English
publishDate 2018
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-61056122018-08-27 Optical Characterization of Neurosurgical Operating Microscopes: Quantitative Fluorescence and Assessment of PpIX Photobleaching Belykh, Evgenii Miller, Eric J. Patel, Arpan A. Bozkurt, Baran Yağmurlu, Kaan Robinson, Timothy R. Nakaji, Peter Spetzler, Robert F. Lawton, Michael T. Nelson, Leonard Y. Seibel, Eric J. Preul, Mark C. Sci Rep Article Protoporphyrin IX (PpIX) induced by 5-aminolevulinic acid (5-ALA) is increasingly used as a fluorescent marker for fluorescence-guided resection of malignant gliomas. Understanding how the properties of the excitation light source and PpIX fluorescence interact with the surgical microscope is critical for effective use of the fluorescence-guided tumor resection technique. In this study, we performed a detailed assessment of the intensity of the emitted blue light and white light and the light beam profile of clinical grade operating microscopes used for PpIX visualization. These measurements revealed both recognized fluorescence photobleaching limitations and unrecognized limitations that may alter quantitative observations of PpIX fluorescence obtained with the operating microscope with potential impact on research and clinical uses. We also evaluated the optical properties of a photostable fluorescent standard with an excitation-emission profile similar to PpIX. In addition, we measured the time-dependent dynamics of 5-ALA-induced PpIX fluorescence in an animal glioma model. Finally, we developed a ratiometric method for quantification of the PpIX fluorescence that uses the photostable fluorescent standard to normalize PpIX fluorescence intensity. This method increases accuracy and allows reproducible and direct comparability of the measurements from multiple samples. Nature Publishing Group UK 2018-08-22 /pmc/articles/PMC6105612/ /pubmed/30135440 http://dx.doi.org/10.1038/s41598-018-30247-6 Text en © The Author(s) 2018 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Belykh, Evgenii
Miller, Eric J.
Patel, Arpan A.
Bozkurt, Baran
Yağmurlu, Kaan
Robinson, Timothy R.
Nakaji, Peter
Spetzler, Robert F.
Lawton, Michael T.
Nelson, Leonard Y.
Seibel, Eric J.
Preul, Mark C.
Optical Characterization of Neurosurgical Operating Microscopes: Quantitative Fluorescence and Assessment of PpIX Photobleaching
title Optical Characterization of Neurosurgical Operating Microscopes: Quantitative Fluorescence and Assessment of PpIX Photobleaching
title_full Optical Characterization of Neurosurgical Operating Microscopes: Quantitative Fluorescence and Assessment of PpIX Photobleaching
title_fullStr Optical Characterization of Neurosurgical Operating Microscopes: Quantitative Fluorescence and Assessment of PpIX Photobleaching
title_full_unstemmed Optical Characterization of Neurosurgical Operating Microscopes: Quantitative Fluorescence and Assessment of PpIX Photobleaching
title_short Optical Characterization of Neurosurgical Operating Microscopes: Quantitative Fluorescence and Assessment of PpIX Photobleaching
title_sort optical characterization of neurosurgical operating microscopes: quantitative fluorescence and assessment of ppix photobleaching
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6105612/
https://www.ncbi.nlm.nih.gov/pubmed/30135440
http://dx.doi.org/10.1038/s41598-018-30247-6
work_keys_str_mv AT belykhevgenii opticalcharacterizationofneurosurgicaloperatingmicroscopesquantitativefluorescenceandassessmentofppixphotobleaching
AT millerericj opticalcharacterizationofneurosurgicaloperatingmicroscopesquantitativefluorescenceandassessmentofppixphotobleaching
AT patelarpana opticalcharacterizationofneurosurgicaloperatingmicroscopesquantitativefluorescenceandassessmentofppixphotobleaching
AT bozkurtbaran opticalcharacterizationofneurosurgicaloperatingmicroscopesquantitativefluorescenceandassessmentofppixphotobleaching
AT yagmurlukaan opticalcharacterizationofneurosurgicaloperatingmicroscopesquantitativefluorescenceandassessmentofppixphotobleaching
AT robinsontimothyr opticalcharacterizationofneurosurgicaloperatingmicroscopesquantitativefluorescenceandassessmentofppixphotobleaching
AT nakajipeter opticalcharacterizationofneurosurgicaloperatingmicroscopesquantitativefluorescenceandassessmentofppixphotobleaching
AT spetzlerrobertf opticalcharacterizationofneurosurgicaloperatingmicroscopesquantitativefluorescenceandassessmentofppixphotobleaching
AT lawtonmichaelt opticalcharacterizationofneurosurgicaloperatingmicroscopesquantitativefluorescenceandassessmentofppixphotobleaching
AT nelsonleonardy opticalcharacterizationofneurosurgicaloperatingmicroscopesquantitativefluorescenceandassessmentofppixphotobleaching
AT seibelericj opticalcharacterizationofneurosurgicaloperatingmicroscopesquantitativefluorescenceandassessmentofppixphotobleaching
AT preulmarkc opticalcharacterizationofneurosurgicaloperatingmicroscopesquantitativefluorescenceandassessmentofppixphotobleaching