Ex vivo , in situ perfusion protocol for human brain fixation compatible with microscopy, MRI techniques, and anatomical studies

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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Bibliographic Details
Published in:Frontiers in neuroanatomy Vol. 17; p. 1149674
Main Authors: 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
Format: Journal Article
Language:English
Published: Switzerland Frontiers Research Foundation 23-03-2023
Frontiers Media S.A
Subjects:
Autopsies
Brain research
carotid perfusion
Clinical Medicine
Donations
Electron microscopy
Ethics
fixation
Histology
human brain
Immunohistochemistry
Intracellular
intracellular injection
Klinisk medicin
Magnetic resonance imaging
Medical and Health Sciences
Medical imaging
Medical research
Medicin och hälsovetenskap
Microscopy
Morphology
MRI
Neuroimaging
Neurological diseases
Neuropathology
Neuroscience
Perfusion
Radiologi och bildbehandling
Radiology, Nuclear Medicine and Medical Imaging
Skull
Spain
carotid perfusion
human brain
histology
MRI
intracellular injection
fixation
electron microscopy
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Description
Summary: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, , 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 , fixation, this procedure can probably be considered the most suitable one available for 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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Reviewed by: Miguel Ángel García-Cabezas, Autonomous University of Madrid, Spain; Alejandra Sierra, University of Eastern Finland, Finland
Edited by: Basilis Zikopoulos, Boston University, United States
ISSN:1662-5129
1662-5129
DOI:10.3389/fnana.2023.1149674