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Analysis of institutional authors

Benavides-Piccione RAuthorTapia-González SAuthor

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May 1, 2023
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Article

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

Publicated to: Frontiers In Neuroanatomy. 17 1149674- - 2023-03-23 17(), DOI: 10.3389/fnana.2023.1149674

Authors: Insausti, R; Insausti, AM; López, MM; Lorenzo, IM; Arroyo-Jiménez, MD; Rabal, MPM; de la Rosa-prieto, C; Delgado-González, JC; Etxeberria, JM; Cebada-Sánchez, S; Raspeño-García, JF; de Onzoño, MMI; Romero, FJM; Benavides-Piccione, R; Tapia-González, S; Wisse, LEM; Ravikumar, S; Wolk, DA; DeFelipe, J; Yushkevich, P; Artacho-Pérula, E

Affiliations

CENTRO DE TECNOLOGIA BIOMEDICA (CTB). Universidad Politécnica de Madrid - Author
CSIC, Inst Cajal, Madrid, Spain - Author
Lund Univ, Dept Diagnost Radiol, Lund, Sweden - Author
Lunds universitet - Author
Penn Medicine - Author
Univ Castilla La Mancha, CRIB, Albacete, Spain - Author
Univ Castilla La Mancha, Sch Med, Med Sci Dept, Neuromax CSIC Associated Unit,Human Neuroanat Lab, Albacete, Spain - Author
Univ Penn, Dept Neurol, Philadelphia, PA USA - Author
Univ Penn, Dept Radiol, Philadelphia, PA USA - Author
Univ Politecn Madrid, Ctr Tecnol Biomed, Lab Cajal Circuitos Cort, Madrid, Spain - Author
Univ Publ Navarra, Sch Med, Dept Hlth, Pamplona, Spain - Author
Universidad de Castilla-La Mancha - Author
Universidad Pública de Navarra - Author
University of Pennsylvania - Author
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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 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.

Keywords

alzheimers-diseasecarotid perfusionchandelier cellselectronelectron microscopyentorhinal cortexfixationhistologyintracellular injectionmonkeymriorganizationparahippocampal subregionspostmortem changestemporal-lobeCarotid perfusionElectron microscopyFixationHistologyHuman brainHuman hippocampal-formationIntracellular injectionMri

Quality index

Bibliometric impact. Analysis of the contribution and dissemination channel

The work has been published in the journal Frontiers In Neuroanatomy due to its progression and the good impact it has achieved in recent years, according to the agency WoS (JCR), it has become a reference in its field. In the year of publication of the work, 2023, it was in position 5/22, thus managing to position itself as a Q1 (Primer Cuartil), in the category Anatomy & Morphology.

From a relative perspective, and based on the normalized impact indicator calculated from World Citations provided by WoS (ESI, Clarivate), it yields a value for the citation normalization relative to the expected citation rate of: 5.5. This indicates that, compared to works in the same discipline and in the same year of publication, it ranks as a work cited above average. (source consulted: ESI Nov 13, 2025)

This information is reinforced by other indicators of the same type, which, although dynamic over time and dependent on the set of average global citations at the time of their calculation, consistently position the work at some point among the top 50% most cited in its field:

  • Weighted Average of Normalized Impact by the Scopus agency: 3.93 (source consulted: FECYT Mar 2025)

Specifically, and according to different indexing agencies, this work has accumulated citations as of 2025-12-05, the following number of citations:

  • WoS: 11
  • Scopus: 11

Impact and social visibility

From the perspective of influence or social adoption, and based on metrics associated with mentions and interactions provided by agencies specializing in calculating the so-called "Alternative or Social Metrics," we can highlight as of 2025-12-05:

  • The use, from an academic perspective evidenced by the Altmetric agency indicator referring to aggregations made by the personal bibliographic manager Mendeley, gives us a total of: 25.
  • The use of this contribution in bookmarks, code forks, additions to favorite lists for recurrent reading, as well as general views, indicates that someone is using the publication as a basis for their current work. This may be a notable indicator of future more formal and academic citations. This claim is supported by the result of the "Capture" indicator, which yields a total of: 24 (PlumX).

With a more dissemination-oriented intent and targeting more general audiences, we can observe other more global scores such as:

  • The Total Score from Altmetric: 2.
  • The number of mentions on the social network X (formerly Twitter): 4 (Altmetric).

It is essential to present evidence supporting full alignment with institutional principles and guidelines on Open Science and the Conservation and Dissemination of Intellectual Heritage. A clear example of this is:

  • The work has been submitted to a journal whose editorial policy allows open Open Access publication.

Leadership analysis of institutional authors

This work has been carried out with international collaboration, specifically with researchers from: Sweden; United States of America.