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Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition

This protocol describes the procedures necessary to support normal metabolic functions of acute brain slice preparations during the collection of magnetic resonance (MR) microscopy data. While it is possible to perform MR collections on living, excised mammalian tissue, such experiments have traditi...

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Autores principales: Flint, Jeremy J., Menon, Kannan, Hansen, Brian, Forder, John, Blackband, Stephen J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5752427/
https://www.ncbi.nlm.nih.gov/pubmed/29155793
http://dx.doi.org/10.3791/56282
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author Flint, Jeremy J.
Menon, Kannan
Hansen, Brian
Forder, John
Blackband, Stephen J.
author_facet Flint, Jeremy J.
Menon, Kannan
Hansen, Brian
Forder, John
Blackband, Stephen J.
author_sort Flint, Jeremy J.
collection PubMed
description This protocol describes the procedures necessary to support normal metabolic functions of acute brain slice preparations during the collection of magnetic resonance (MR) microscopy data. While it is possible to perform MR collections on living, excised mammalian tissue, such experiments have traditionally been constrained by resolution limits and are thus incapable of visualizing tissue microstructure. Conversely, MR protocols that did achieve microscopic image resolution required the use of fixed samples to accommodate the need for static, unchanging conditions over lengthy scan times. The current protocol describes the first available MR technique that enables imaging of living, mammalian tissue samples at microscopic resolutions. Such data is of great importance to the understanding of how pathology-based contrast changes occurring at the microscopic level influence the content of macroscopic MR scans such as those used in the clinic. Once such an understanding is realized, diagnostic methods with greater sensitivity and accuracy can be developed, which will translate directly to earlier disease treatment, more accurate therapy monitoring and improved patient outcomes. While the described methodology focuses on brain slice preparations, the protocol is adaptable to any excised tissue slice given that changes are made to the gas and perfusate preparations to accommodate the tissue's specific metabolic needs. Successful execution of the protocol should result in living, acute slice preparations that exhibit MR diffusion signal stability for periods up to 15.5 h. The primary advantages of the current system over other MR compatible perfusion apparatuses are its compatibility with the MR microscopy hardware required to attain higher resolution images and ability to provide constant, uninterrupted flow with carefully regulated perfusate conditions. Reduced sample throughput is a consideration with this design as only one tissue slice may be imaged at a time.
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spelling pubmed-57524272018-01-19 Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition Flint, Jeremy J. Menon, Kannan Hansen, Brian Forder, John Blackband, Stephen J. J Vis Exp Bioengineering This protocol describes the procedures necessary to support normal metabolic functions of acute brain slice preparations during the collection of magnetic resonance (MR) microscopy data. While it is possible to perform MR collections on living, excised mammalian tissue, such experiments have traditionally been constrained by resolution limits and are thus incapable of visualizing tissue microstructure. Conversely, MR protocols that did achieve microscopic image resolution required the use of fixed samples to accommodate the need for static, unchanging conditions over lengthy scan times. The current protocol describes the first available MR technique that enables imaging of living, mammalian tissue samples at microscopic resolutions. Such data is of great importance to the understanding of how pathology-based contrast changes occurring at the microscopic level influence the content of macroscopic MR scans such as those used in the clinic. Once such an understanding is realized, diagnostic methods with greater sensitivity and accuracy can be developed, which will translate directly to earlier disease treatment, more accurate therapy monitoring and improved patient outcomes. While the described methodology focuses on brain slice preparations, the protocol is adaptable to any excised tissue slice given that changes are made to the gas and perfusate preparations to accommodate the tissue's specific metabolic needs. Successful execution of the protocol should result in living, acute slice preparations that exhibit MR diffusion signal stability for periods up to 15.5 h. The primary advantages of the current system over other MR compatible perfusion apparatuses are its compatibility with the MR microscopy hardware required to attain higher resolution images and ability to provide constant, uninterrupted flow with carefully regulated perfusate conditions. Reduced sample throughput is a consideration with this design as only one tissue slice may be imaged at a time. MyJove Corporation 2017-10-18 /pmc/articles/PMC5752427/ /pubmed/29155793 http://dx.doi.org/10.3791/56282 Text en Copyright © 2017, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/
spellingShingle Bioengineering
Flint, Jeremy J.
Menon, Kannan
Hansen, Brian
Forder, John
Blackband, Stephen J.
Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition
title Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition
title_full Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition
title_fullStr Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition
title_full_unstemmed Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition
title_short Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition
title_sort metabolic support of excised, living brain tissues during magnetic resonance microscopy acquisition
topic Bioengineering
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5752427/
https://www.ncbi.nlm.nih.gov/pubmed/29155793
http://dx.doi.org/10.3791/56282
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