Autofluorescence enhancement for label-free imaging of myelinated fibers in mammalian brains

Abstract Analyzing the structure of neuronal fibers with single axon resolution in large volumes is a challenge in connectomics. Different technologies try to address this goal; however, they are limited either by the ineffective labeling of the fibers or in the achievable resolution. The possibilit...

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Autores principales:	Irene Costantini, Enrico Baria, Michele Sorelli, Felix Matuschke, Francesco Giardini, Miriam Menzel, Giacomo Mazzamuto, Ludovico Silvestri, Riccardo Cicchi, Katrin Amunts, Markus Axer, Francesco Saverio Pavone
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/2c8d7874d29b4aaea0902cccf378eba9
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spelling	oai:doaj.org-article:2c8d7874d29b4aaea0902cccf378eba92021-12-02T15:51:12ZAutofluorescence enhancement for label-free imaging of myelinated fibers in mammalian brains10.1038/s41598-021-86092-72045-2322https://doaj.org/article/2c8d7874d29b4aaea0902cccf378eba92021-04-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-86092-7https://doaj.org/toc/2045-2322Abstract Analyzing the structure of neuronal fibers with single axon resolution in large volumes is a challenge in connectomics. Different technologies try to address this goal; however, they are limited either by the ineffective labeling of the fibers or in the achievable resolution. The possibility of discriminating between different adjacent myelinated axons gives the opportunity of providing more information about the fiber composition and architecture within a specific area. Here, we propose MAGIC (Myelin Autofluorescence imaging by Glycerol Induced Contrast enhancement), a tissue preparation method to perform label-free fluorescence imaging of myelinated fibers that is user friendly and easy to handle. We exploit the high axial and radial resolution of two-photon fluorescence microscopy (TPFM) optical sectioning to decipher the mixture of various fiber orientations within the sample of interest. We demonstrate its broad applicability by performing mesoscopic reconstruction at a sub-micron resolution of mouse, rat, monkey, and human brain samples and by quantifying the different fiber organization in control and Reeler mouse's hippocampal sections. Our study provides a novel method for 3D label-free imaging of nerve fibers in fixed samples at high resolution, below micrometer level, that overcomes the limitation related to the myelinated axons exogenous labeling, improving the possibility of analyzing brain connectivity.Irene CostantiniEnrico BariaMichele SorelliFelix MatuschkeFrancesco GiardiniMiriam MenzelGiacomo MazzamutoLudovico SilvestriRiccardo CicchiKatrin AmuntsMarkus AxerFrancesco Saverio PavoneNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-11 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Irene Costantini Enrico Baria Michele Sorelli Felix Matuschke Francesco Giardini Miriam Menzel Giacomo Mazzamuto Ludovico Silvestri Riccardo Cicchi Katrin Amunts Markus Axer Francesco Saverio Pavone Autofluorescence enhancement for label-free imaging of myelinated fibers in mammalian brains
description	Abstract Analyzing the structure of neuronal fibers with single axon resolution in large volumes is a challenge in connectomics. Different technologies try to address this goal; however, they are limited either by the ineffective labeling of the fibers or in the achievable resolution. The possibility of discriminating between different adjacent myelinated axons gives the opportunity of providing more information about the fiber composition and architecture within a specific area. Here, we propose MAGIC (Myelin Autofluorescence imaging by Glycerol Induced Contrast enhancement), a tissue preparation method to perform label-free fluorescence imaging of myelinated fibers that is user friendly and easy to handle. We exploit the high axial and radial resolution of two-photon fluorescence microscopy (TPFM) optical sectioning to decipher the mixture of various fiber orientations within the sample of interest. We demonstrate its broad applicability by performing mesoscopic reconstruction at a sub-micron resolution of mouse, rat, monkey, and human brain samples and by quantifying the different fiber organization in control and Reeler mouse's hippocampal sections. Our study provides a novel method for 3D label-free imaging of nerve fibers in fixed samples at high resolution, below micrometer level, that overcomes the limitation related to the myelinated axons exogenous labeling, improving the possibility of analyzing brain connectivity.
format	article
author	Irene Costantini Enrico Baria Michele Sorelli Felix Matuschke Francesco Giardini Miriam Menzel Giacomo Mazzamuto Ludovico Silvestri Riccardo Cicchi Katrin Amunts Markus Axer Francesco Saverio Pavone
author_facet	Irene Costantini Enrico Baria Michele Sorelli Felix Matuschke Francesco Giardini Miriam Menzel Giacomo Mazzamuto Ludovico Silvestri Riccardo Cicchi Katrin Amunts Markus Axer Francesco Saverio Pavone
author_sort	Irene Costantini
title	Autofluorescence enhancement for label-free imaging of myelinated fibers in mammalian brains
title_short	Autofluorescence enhancement for label-free imaging of myelinated fibers in mammalian brains
title_full	Autofluorescence enhancement for label-free imaging of myelinated fibers in mammalian brains
title_fullStr	Autofluorescence enhancement for label-free imaging of myelinated fibers in mammalian brains
title_full_unstemmed	Autofluorescence enhancement for label-free imaging of myelinated fibers in mammalian brains
title_sort	autofluorescence enhancement for label-free imaging of myelinated fibers in mammalian brains
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/2c8d7874d29b4aaea0902cccf378eba9
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_version_	1718385621722988544

Autofluorescence enhancement for label-free imaging of myelinated fibers in mammalian brains

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