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Ex vivo, in situ perfusion protocol for human brain fixation compatible with microscopy, MRI techniques, and anatomical studies
We present a method for human brain fixation based on simultaneous perfusion of 4% paraformaldehyde through carotids after a flush with saline. The left carotid cannula is used to perfuse the body with 10% formalin, to allow further use of the body for anatomical research or teaching. The aim of our...
Autores principales: | , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10076536/ https://www.ncbi.nlm.nih.gov/pubmed/37034833 http://dx.doi.org/10.3389/fnana.2023.1149674 |
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author | Insausti, Ricardo Insausti, Ana María Muñoz López, Mónica Medina Lorenzo, Isidro Arroyo-Jiménez, Maria del Mar Marcos Rabal, María Pilar de la Rosa-Prieto, Carlos Delgado-González, José Carlos Montón Etxeberria, Javier Cebada-Sánchez, Sandra Raspeño-García, Juan Francisco Iñiguez de Onzoño, María Mercedes Molina Romero, Francisco Javier Benavides-Piccione, Ruth Tapia-González, Silvia Wisse, Laura E. M. Ravikumar, Sadhana Wolk, David A. DeFelipe, Javier Yushkevich, Paul Artacho-Pérula, Emilio |
author_facet | Insausti, Ricardo Insausti, Ana María Muñoz López, Mónica Medina Lorenzo, Isidro Arroyo-Jiménez, Maria del Mar Marcos Rabal, María Pilar de la Rosa-Prieto, Carlos Delgado-González, José Carlos Montón Etxeberria, Javier Cebada-Sánchez, Sandra Raspeño-García, Juan Francisco Iñiguez de Onzoño, María Mercedes Molina Romero, Francisco Javier Benavides-Piccione, Ruth Tapia-González, Silvia Wisse, Laura E. M. Ravikumar, Sadhana Wolk, David A. DeFelipe, Javier Yushkevich, Paul Artacho-Pérula, Emilio |
author_sort | Insausti, Ricardo |
collection | PubMed |
description | We present a method for human brain fixation based on simultaneous perfusion of 4% paraformaldehyde through carotids after a flush with saline. The left carotid cannula is used to perfuse the body with 10% formalin, to allow further use of the body for anatomical research or teaching. The aim of our method is to develop a vascular fixation protocol for the human brain, by adapting protocols that are commonly used in experimental animal studies. We show that a variety of histological procedures can be carried out (cyto- and myeloarchitectonics, histochemistry, immunohistochemistry, intracellular cell injection, and electron microscopy). In addition, ex vivo, ex situ high-resolution MRI (9.4T) can be obtained in the same specimens. This procedure resulted in similar morphological features to those obtained by intravascular perfusion in experimental animals, provided that the postmortem interval was under 10 h for several of the techniques used and under 4 h in the case of intracellular injections and electron microscopy. The use of intravascular fixation of the brain inside the skull provides a fixed whole human brain, perfectly fitted to the skull, with negligible deformation compared to conventional techniques. Given this characteristic of ex vivo, in situ fixation, this procedure can probably be considered the most suitable one available for ex vivo MRI scans of the brain. We describe the compatibility of the method proposed for intravascular fixation of the human brain and fixation of the donor’s body for anatomical purposes. Thus, body donor programs can provide human brain tissue, while the remainder of the body can also be fixed for anatomical studies. Therefore, this method of human brain fixation through the carotid system optimizes the procurement of human brain tissue, allowing a greater understanding of human neurological diseases, while benefiting anatomy departments by making the remainder of the body available for teaching purposes. |
format | Online Article Text |
id | pubmed-10076536 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-100765362023-04-07 Ex vivo, in situ perfusion protocol for human brain fixation compatible with microscopy, MRI techniques, and anatomical studies Insausti, Ricardo Insausti, Ana María Muñoz López, Mónica Medina Lorenzo, Isidro Arroyo-Jiménez, Maria del Mar Marcos Rabal, María Pilar de la Rosa-Prieto, Carlos Delgado-González, José Carlos Montón Etxeberria, Javier Cebada-Sánchez, Sandra Raspeño-García, Juan Francisco Iñiguez de Onzoño, María Mercedes Molina Romero, Francisco Javier Benavides-Piccione, Ruth Tapia-González, Silvia Wisse, Laura E. M. Ravikumar, Sadhana Wolk, David A. DeFelipe, Javier Yushkevich, Paul Artacho-Pérula, Emilio Front Neuroanat Neuroscience We present a method for human brain fixation based on simultaneous perfusion of 4% paraformaldehyde through carotids after a flush with saline. The left carotid cannula is used to perfuse the body with 10% formalin, to allow further use of the body for anatomical research or teaching. The aim of our method is to develop a vascular fixation protocol for the human brain, by adapting protocols that are commonly used in experimental animal studies. We show that a variety of histological procedures can be carried out (cyto- and myeloarchitectonics, histochemistry, immunohistochemistry, intracellular cell injection, and electron microscopy). In addition, ex vivo, ex situ high-resolution MRI (9.4T) can be obtained in the same specimens. This procedure resulted in similar morphological features to those obtained by intravascular perfusion in experimental animals, provided that the postmortem interval was under 10 h for several of the techniques used and under 4 h in the case of intracellular injections and electron microscopy. The use of intravascular fixation of the brain inside the skull provides a fixed whole human brain, perfectly fitted to the skull, with negligible deformation compared to conventional techniques. Given this characteristic of ex vivo, in situ fixation, this procedure can probably be considered the most suitable one available for ex vivo MRI scans of the brain. We describe the compatibility of the method proposed for intravascular fixation of the human brain and fixation of the donor’s body for anatomical purposes. Thus, body donor programs can provide human brain tissue, while the remainder of the body can also be fixed for anatomical studies. Therefore, this method of human brain fixation through the carotid system optimizes the procurement of human brain tissue, allowing a greater understanding of human neurological diseases, while benefiting anatomy departments by making the remainder of the body available for teaching purposes. Frontiers Media S.A. 2023-03-23 /pmc/articles/PMC10076536/ /pubmed/37034833 http://dx.doi.org/10.3389/fnana.2023.1149674 Text en Copyright © 2023 Insausti, Insausti, Muñoz López, Medina Lorenzo, Arroyo-Jiménez, Marcos Rabal, de La Rosa-Prieto, Delgado-González, Montón Etxeberria, Cebada-Sánchez, Raspeño-García, Iñiguez de Onzoño, Molina Romero, Benavides-Piccione, Tapia-González, Wisse, Ravikumar, Wolk, DeFelipe, Yushkevich and Artacho-Pérula. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Neuroscience Insausti, Ricardo Insausti, Ana María Muñoz López, Mónica Medina Lorenzo, Isidro Arroyo-Jiménez, Maria del Mar Marcos Rabal, María Pilar de la Rosa-Prieto, Carlos Delgado-González, José Carlos Montón Etxeberria, Javier Cebada-Sánchez, Sandra Raspeño-García, Juan Francisco Iñiguez de Onzoño, María Mercedes Molina Romero, Francisco Javier Benavides-Piccione, Ruth Tapia-González, Silvia Wisse, Laura E. M. Ravikumar, Sadhana Wolk, David A. DeFelipe, Javier Yushkevich, Paul Artacho-Pérula, Emilio Ex vivo, in situ perfusion protocol for human brain fixation compatible with microscopy, MRI techniques, and anatomical studies |
title | Ex vivo, in situ perfusion protocol for human brain fixation compatible with microscopy, MRI techniques, and anatomical studies |
title_full | Ex vivo, in situ perfusion protocol for human brain fixation compatible with microscopy, MRI techniques, and anatomical studies |
title_fullStr | Ex vivo, in situ perfusion protocol for human brain fixation compatible with microscopy, MRI techniques, and anatomical studies |
title_full_unstemmed | Ex vivo, in situ perfusion protocol for human brain fixation compatible with microscopy, MRI techniques, and anatomical studies |
title_short | Ex vivo, in situ perfusion protocol for human brain fixation compatible with microscopy, MRI techniques, and anatomical studies |
title_sort | ex vivo, in situ perfusion protocol for human brain fixation compatible with microscopy, mri techniques, and anatomical studies |
topic | Neuroscience |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10076536/ https://www.ncbi.nlm.nih.gov/pubmed/37034833 http://dx.doi.org/10.3389/fnana.2023.1149674 |
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