Comparison of Different Tissue Clearing Methods for Three-Dimensional Reconstruction of Human Brain Cellular Anatomy Using Advanced Imaging Techniques

The combination of tissue clearing techniques with advanced optical microscopy facilitates the achievement of three-dimensional (3D) reconstruction of macroscopic specimens at high resolution. Whole mouse organs or even bodies have been analyzed, while the reconstruction of the human nervous system...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Marina Scardigli, Luca Pesce, Niamh Brady, Giacomo Mazzamuto, Vladislav Gavryusev, Ludovico Silvestri, Patrick R. Hof, Christophe Destrieux, Irene Costantini, Francesco S. Pavone
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	clearing techniques optical microscopy immunofluorescence light-sheet fluorescence microscopy expansion microscopy Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571 Human anatomy QM1-695
Acceso en línea:	https://doaj.org/article/7792613291ca417ab4f5bdea08653169
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:7792613291ca417ab4f5bdea08653169
record_format	dspace
spelling	oai:doaj.org-article:7792613291ca417ab4f5bdea086531692021-11-11T14:16:23ZComparison of Different Tissue Clearing Methods for Three-Dimensional Reconstruction of Human Brain Cellular Anatomy Using Advanced Imaging Techniques1662-512910.3389/fnana.2021.752234https://doaj.org/article/7792613291ca417ab4f5bdea086531692021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fnana.2021.752234/fullhttps://doaj.org/toc/1662-5129The combination of tissue clearing techniques with advanced optical microscopy facilitates the achievement of three-dimensional (3D) reconstruction of macroscopic specimens at high resolution. Whole mouse organs or even bodies have been analyzed, while the reconstruction of the human nervous system remains a challenge. Although several tissue protocols have been proposed, the high autofluorescence and variable post-mortem conditions of human specimens negatively affect the quality of the images in terms of achievable transparency and staining contrast. Moreover, homogeneous staining of high-density epitopes, such as neuronal nuclear antigen (NeuN), creates an additional challenge. Here, we evaluated different tissue transformation approaches to find the best solution to uniformly clear and label all neurons in the human cerebral cortex using anti-NeuN antibodies in combination with confocal and light-sheet fluorescence microscopy (LSFM). Finally, we performed mesoscopic high-resolution 3D reconstruction of the successfully clarified and stained samples with LSFM.Marina ScardigliMarina ScardigliLuca PesceLuca PesceNiamh BradyGiacomo MazzamutoGiacomo MazzamutoVladislav GavryusevVladislav GavryusevLudovico SilvestriLudovico SilvestriLudovico SilvestriPatrick R. HofChristophe DestrieuxIrene CostantiniIrene CostantiniIrene CostantiniFrancesco S. PavoneFrancesco S. PavoneFrancesco S. PavoneFrontiers Media S.A.articleclearing techniquesoptical microscopyimmunofluorescencelight-sheet fluorescence microscopyexpansion microscopyNeurosciences. Biological psychiatry. NeuropsychiatryRC321-571Human anatomyQM1-695ENFrontiers in Neuroanatomy, Vol 15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	clearing techniques optical microscopy immunofluorescence light-sheet fluorescence microscopy expansion microscopy Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571 Human anatomy QM1-695
spellingShingle	clearing techniques optical microscopy immunofluorescence light-sheet fluorescence microscopy expansion microscopy Neurosciences. Biological psychiatry. Neuropsychiatry RC321-571 Human anatomy QM1-695 Marina Scardigli Marina Scardigli Luca Pesce Luca Pesce Niamh Brady Giacomo Mazzamuto Giacomo Mazzamuto Vladislav Gavryusev Vladislav Gavryusev Ludovico Silvestri Ludovico Silvestri Ludovico Silvestri Patrick R. Hof Christophe Destrieux Irene Costantini Irene Costantini Irene Costantini Francesco S. Pavone Francesco S. Pavone Francesco S. Pavone Comparison of Different Tissue Clearing Methods for Three-Dimensional Reconstruction of Human Brain Cellular Anatomy Using Advanced Imaging Techniques
description	The combination of tissue clearing techniques with advanced optical microscopy facilitates the achievement of three-dimensional (3D) reconstruction of macroscopic specimens at high resolution. Whole mouse organs or even bodies have been analyzed, while the reconstruction of the human nervous system remains a challenge. Although several tissue protocols have been proposed, the high autofluorescence and variable post-mortem conditions of human specimens negatively affect the quality of the images in terms of achievable transparency and staining contrast. Moreover, homogeneous staining of high-density epitopes, such as neuronal nuclear antigen (NeuN), creates an additional challenge. Here, we evaluated different tissue transformation approaches to find the best solution to uniformly clear and label all neurons in the human cerebral cortex using anti-NeuN antibodies in combination with confocal and light-sheet fluorescence microscopy (LSFM). Finally, we performed mesoscopic high-resolution 3D reconstruction of the successfully clarified and stained samples with LSFM.
format	article
author	Marina Scardigli Marina Scardigli Luca Pesce Luca Pesce Niamh Brady Giacomo Mazzamuto Giacomo Mazzamuto Vladislav Gavryusev Vladislav Gavryusev Ludovico Silvestri Ludovico Silvestri Ludovico Silvestri Patrick R. Hof Christophe Destrieux Irene Costantini Irene Costantini Irene Costantini Francesco S. Pavone Francesco S. Pavone Francesco S. Pavone
author_facet	Marina Scardigli Marina Scardigli Luca Pesce Luca Pesce Niamh Brady Giacomo Mazzamuto Giacomo Mazzamuto Vladislav Gavryusev Vladislav Gavryusev Ludovico Silvestri Ludovico Silvestri Ludovico Silvestri Patrick R. Hof Christophe Destrieux Irene Costantini Irene Costantini Irene Costantini Francesco S. Pavone Francesco S. Pavone Francesco S. Pavone
author_sort	Marina Scardigli
title	Comparison of Different Tissue Clearing Methods for Three-Dimensional Reconstruction of Human Brain Cellular Anatomy Using Advanced Imaging Techniques
title_short	Comparison of Different Tissue Clearing Methods for Three-Dimensional Reconstruction of Human Brain Cellular Anatomy Using Advanced Imaging Techniques
title_full	Comparison of Different Tissue Clearing Methods for Three-Dimensional Reconstruction of Human Brain Cellular Anatomy Using Advanced Imaging Techniques
title_fullStr	Comparison of Different Tissue Clearing Methods for Three-Dimensional Reconstruction of Human Brain Cellular Anatomy Using Advanced Imaging Techniques
title_full_unstemmed	Comparison of Different Tissue Clearing Methods for Three-Dimensional Reconstruction of Human Brain Cellular Anatomy Using Advanced Imaging Techniques
title_sort	comparison of different tissue clearing methods for three-dimensional reconstruction of human brain cellular anatomy using advanced imaging techniques
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/7792613291ca417ab4f5bdea08653169
work_keys_str_mv	AT marinascardigli comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT marinascardigli comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT lucapesce comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT lucapesce comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT niamhbrady comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT giacomomazzamuto comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT giacomomazzamuto comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT vladislavgavryusev comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT vladislavgavryusev comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT ludovicosilvestri comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT ludovicosilvestri comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT ludovicosilvestri comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT patrickrhof comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT christophedestrieux comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT irenecostantini comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT irenecostantini comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT irenecostantini comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT francescospavone comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT francescospavone comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques AT francescospavone comparisonofdifferenttissueclearingmethodsforthreedimensionalreconstructionofhumanbraincellularanatomyusingadvancedimagingtechniques
_version_	1718439047450329088

Comparison of Different Tissue Clearing Methods for Three-Dimensional Reconstruction of Human Brain Cellular Anatomy Using Advanced Imaging Techniques

Ejemplares similares