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
Human Neuroanatomy Laboratory, Neuromax CSIC Associated Unit, Medical Sciences Department, School of Medicine and CRIB, University of Castilla La Mancha, Albacete, Spain.
Department of Health, School of Medicine, Public University of Navarra, Pamplona, Spain.
Front Neuroanat. 2023 Mar 23;17:1149674. doi: 10.3389/fnana.2023.1149674. eCollection 2023.
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.
我们提出了一种人脑固定方法,即在先用生理盐水冲洗后,通过颈动脉同时灌注4%的多聚甲醛。左颈动脉插管用于向身体灌注10%的福尔马林,以便将身体进一步用于解剖学研究或教学。我们方法的目的是通过改编实验动物研究中常用的方案,开发一种用于人脑的血管固定方案。我们表明,可以进行多种组织学程序(细胞和髓质结构学、组织化学、免疫组织化学、细胞内注射和电子显微镜检查)。此外,还可以在相同的标本上获得高分辨率MRI(9.4T)。如果使用的几种技术的死后间隔在10小时以内,细胞内注射和电子显微镜检查的死后间隔在4小时以内,该程序会产生与实验动物血管内灌注所获得的形态特征相似的结果。在颅骨内对脑进行血管内固定,可提供一个与颅骨完美贴合的固定全脑,与传统技术相比变形可忽略不计。鉴于这种固定的特性,该程序可能被认为是最适合用于脑部MRI扫描的方法。我们描述了所提出的人脑血管内固定方法与为解剖目的固定供体身体的兼容性。因此,尸体捐赠项目可以提供人脑组织,而身体的其余部分也可以固定用于解剖学研究。因此,这种通过颈动脉系统进行人脑固定的方法优化了人脑组织的获取,有助于更深入地了解人类神经疾病,同时通过使身体的其余部分可用于教学目的而使解剖学系受益。