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New Design of the Electrophoretic Part of CLARITY Technology for Confocal Light Microscopy of Rat and Human Brains

Background: CLARITY is a method of rendering postmortem brain tissue transparent using acrylamide-based hydrogels so that this tissue could be further used for immunohistochemistry, molecular biology, or gross anatomical studies. Published papers using the CLARITY method have included studies on hum...

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Autores principales: Zach, Petr, Mrzílková, Jana, Pala, Jan, Uttl, Libor, Kútna, Viera, Musil, Vladimír, Sommerová, Blanka, Tůma, Petr
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6770398/
https://www.ncbi.nlm.nih.gov/pubmed/31470513
http://dx.doi.org/10.3390/brainsci9090218
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author Zach, Petr
Mrzílková, Jana
Pala, Jan
Uttl, Libor
Kútna, Viera
Musil, Vladimír
Sommerová, Blanka
Tůma, Petr
author_facet Zach, Petr
Mrzílková, Jana
Pala, Jan
Uttl, Libor
Kútna, Viera
Musil, Vladimír
Sommerová, Blanka
Tůma, Petr
author_sort Zach, Petr
collection PubMed
description Background: CLARITY is a method of rendering postmortem brain tissue transparent using acrylamide-based hydrogels so that this tissue could be further used for immunohistochemistry, molecular biology, or gross anatomical studies. Published papers using the CLARITY method have included studies on human brains suffering from Alzheimer’s disease using mouse spinal cords as animal models for multiple sclerosis. Methods: We modified the original design of the Chung CLARITY system by altering the electrophoretic flow-through cell, the shape of the platinum electrophoresis electrodes and their positions, as well as the cooling and recirculation system, so that it provided a greater effect and can be used in any laboratory. Results: The adapted CLARITY system is assembled from basic laboratory components, in contrast to the original design. The modified CLARITY system was tested both on rat brain stained with a rabbit polyclonal anti-Iba-1 for microglial cells and on human nucleus accumbens stained with parvalbumin and tyrosine hydroxylase for visualization of specific neurons by confocal laser scanning microscopy. Conclusions: Our design has the advantage of simplicity, functional robustness, and minimal requirement for specialized additional items for the construction of the CLARITY apparatus.
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spelling pubmed-67703982019-10-30 New Design of the Electrophoretic Part of CLARITY Technology for Confocal Light Microscopy of Rat and Human Brains Zach, Petr Mrzílková, Jana Pala, Jan Uttl, Libor Kútna, Viera Musil, Vladimír Sommerová, Blanka Tůma, Petr Brain Sci Article Background: CLARITY is a method of rendering postmortem brain tissue transparent using acrylamide-based hydrogels so that this tissue could be further used for immunohistochemistry, molecular biology, or gross anatomical studies. Published papers using the CLARITY method have included studies on human brains suffering from Alzheimer’s disease using mouse spinal cords as animal models for multiple sclerosis. Methods: We modified the original design of the Chung CLARITY system by altering the electrophoretic flow-through cell, the shape of the platinum electrophoresis electrodes and their positions, as well as the cooling and recirculation system, so that it provided a greater effect and can be used in any laboratory. Results: The adapted CLARITY system is assembled from basic laboratory components, in contrast to the original design. The modified CLARITY system was tested both on rat brain stained with a rabbit polyclonal anti-Iba-1 for microglial cells and on human nucleus accumbens stained with parvalbumin and tyrosine hydroxylase for visualization of specific neurons by confocal laser scanning microscopy. Conclusions: Our design has the advantage of simplicity, functional robustness, and minimal requirement for specialized additional items for the construction of the CLARITY apparatus. MDPI 2019-08-29 /pmc/articles/PMC6770398/ /pubmed/31470513 http://dx.doi.org/10.3390/brainsci9090218 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zach, Petr
Mrzílková, Jana
Pala, Jan
Uttl, Libor
Kútna, Viera
Musil, Vladimír
Sommerová, Blanka
Tůma, Petr
New Design of the Electrophoretic Part of CLARITY Technology for Confocal Light Microscopy of Rat and Human Brains
title New Design of the Electrophoretic Part of CLARITY Technology for Confocal Light Microscopy of Rat and Human Brains
title_full New Design of the Electrophoretic Part of CLARITY Technology for Confocal Light Microscopy of Rat and Human Brains
title_fullStr New Design of the Electrophoretic Part of CLARITY Technology for Confocal Light Microscopy of Rat and Human Brains
title_full_unstemmed New Design of the Electrophoretic Part of CLARITY Technology for Confocal Light Microscopy of Rat and Human Brains
title_short New Design of the Electrophoretic Part of CLARITY Technology for Confocal Light Microscopy of Rat and Human Brains
title_sort new design of the electrophoretic part of clarity technology for confocal light microscopy of rat and human brains
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6770398/
https://www.ncbi.nlm.nih.gov/pubmed/31470513
http://dx.doi.org/10.3390/brainsci9090218
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