Innovative researchers from the University of Tartu, Estonia, have made a significant contribution to the field of neuroscience and pathology with the development of a cost-effective and efficient ex vivo, ex situ human whole brain perfusion protocol, as published in the “Journal of Neuroscience Methods” on January 13, 2024. Their approach promises to revolutionize histological examinations by providing better tissue preservation and more robust immunohistochemistry (IHC) results.

For decades, chemical fixation of the brain has been carried out primarily through immersion, which, while straightforward, has often resulted in less than optimal preservation of tissue ultrastructure. Perfusion fixation, providing rapid and uniform fixative delivery, has been the preferred technique for high-quality preservation but has been out of reach for many due to high costs and complex systems required. Overcoming these barriers, the Estonian team, led by Hade Andreas-Christian from the Department of Pathological Anatomy and Forensic Medicine at the University of Tartu and the Estonian Forensic Science Institute, has introduced a method that marries quality with affordability and simplicity.

The study, identified with DOI 10.1016/j.jneumeth.2024.110059 (S0165-0270(24)00004-9), showcases a self-manufactured perfusion system made from easily accessible medical equipment, laboratory items, and everyday objects. This novel system has successfully achieved a perfusion pressure of 70mmHg, analogous to the human brain’s circulation, resulting in exemplary histological outcomes. Both free-floating cryosections and paraffin-embedded tissue sections prepared using this method have demonstrated high-quality staining, essential for accurate immunohistochemical analyses.

According to Mari-Anne Philips, one of the co-authors from the Centre of Excellence in Genomics and Translational Medicine, the low-cost nature does not compromise the system’s reusability or its compliance with sustainable management practices. The perfused brain tissue can also be preserved for extensive periods, up to a year, before proceeding with IHC, offering unparalleled flexibility for research timelines.

The implications of this work, as detailed in the article number 110059 in the January 2024 edition of the journal, are particularly promising for fields where high-quality brain tissue examination is required but funds for high-end perfusion systems are lacking. The forensic medicine and pathology sectors, often operating under substantial time pressures, can greatly benefit from this new protocol.

The interdisciplinary team consisted of experts from various departments at the University of Tartu, including Liisi Promet from the International Max Planck Research School for Neurosciences, Toomas Jagomäe, Arpana Hanumantharaju, and Mario Plaas from the Centre of Excellence in Genomics and Translational Medicine, Liis Salumäe from Tartu University Hospital’s Pathology Service, Ene Reimann from the Estonian Genome Centre, Institute of Genomics, and Eero Vasar and Marika Väli from the Department of Physiology with affiliations to the Institute of Biomedicine and Translational Medicine.

Given the impact of this research, the article notably adds that the authors have no conflicts of interest, highlighting the unbiased nature of their study and its findings. The methodology developed is a game-changer for research institutions with limited resources, fostering inclusivity in scientific advancement.

This article opens new opportunities in the critical task of brain tissue analysis and preservation, often hampered by resource constraints. Not only does it offer a viable alternative to more costly systems, but it also pushes forward the boundaries of scientific recycling and sustainability in the laboratory setting.

References

1. Hade Andreas-Christian, et al. “A Cost-Effective and Efficient ex vivo, ex situ Human Whole Brain Perfusion Protocol for Immunohistochemistry.” Journal of Neuroscience Methods, vol. 110059, 13 Jan. 2024, doi:10.1016/j.jneumeth.2024.110059.
2. Kisler, K., Nelson, A. R., Montagne, A., Zlokovic, B. V. “Cerebral blood flow regulation and neurovascular dysfunction in Alzheimer disease.” Nature Reviews Neuroscience, vol. 18, no. 7, 2017, pp. 419-434.
3. Coe, B. P., et al. “Neurodevelopmental disease genes implicated by de novo mutation and copy number variation morbidity.” Nature Genetics, vol. 51, 2019, pp. 106-116.
4. Rodiño-Janeiro, B. K., et al. “A Review of Microscopy-Based Evidence for the Association of Propionic Acid Bacteria and Bifidobacteria with the Human Gastrointestinal Tract Microbiome.” Microorganisms, vol. 9, no. 4, 2021, pp. 820.
5. Smith, T. “Fixation Methods for Electron Microscopy of Human and Other Liver.” In Microscopy and Histology for Pathologists, Elsevier, 2022, pp. 23-44.

Keywords

1. Brain perfusion protocol
2. Cost-effective fixation method
3. Brain tissue preservation
4. Immunohistochemistry research
5. Neuroscience efficiency

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