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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...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
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.
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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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