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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 and Cebada-Sánchez, Sandra , et al. (2023) In Frontiers in Neuroanatomy 17.
Abstract

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... (More)

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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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
carotid perfusion, electron microscopy, fixation, histology, human brain, intracellular injection, MRI
in
Frontiers in Neuroanatomy
volume
17
article number
1149674
publisher
Frontiers Media S. A.
external identifiers
  • pmid:37034833
  • scopus:85153403832
ISSN
1662-5129
DOI
10.3389/fnana.2023.1149674
language
English
LU publication?
yes
id
2f1886b7-17a6-4657-b491-46287b2b9c43
date added to LUP
2023-07-14 10:54:01
date last changed
2024-04-19 23:26:55
@article{2f1886b7-17a6-4657-b491-46287b2b9c43,
  abstract     = {{<p>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.</p>}},
  author       = {{Insausti, Ricardo and Insausti, Ana María and Muñoz López, Mónica and Medina Lorenzo, Isidro and Arroyo-Jiménez, Maria del Mar and Marcos Rabal, María Pilar and de la Rosa-Prieto, Carlos and Delgado-González, José Carlos and Montón Etxeberria, Javier and Cebada-Sánchez, Sandra and Raspeño-García, Juan Francisco and Iñiguez de Onzoño, María Mercedes and Molina Romero, Francisco Javier and Benavides-Piccione, Ruth and Tapia-González, Silvia and Wisse, Laura E.M. and Ravikumar, Sadhana and Wolk, David A. and DeFelipe, Javier and Yushkevich, Paul and Artacho-Pérula, Emilio}},
  issn         = {{1662-5129}},
  keywords     = {{carotid perfusion; electron microscopy; fixation; histology; human brain; intracellular injection; MRI}},
  language     = {{eng}},
  publisher    = {{Frontiers Media S. A.}},
  series       = {{Frontiers in Neuroanatomy}},
  title        = {{Ex vivo, in situ perfusion protocol for human brain fixation compatible with microscopy, MRI techniques, and anatomical studies}},
  url          = {{http://dx.doi.org/10.3389/fnana.2023.1149674}},
  doi          = {{10.3389/fnana.2023.1149674}},
  volume       = {{17}},
  year         = {{2023}},
}