Ex situ perfusion fixation for brain banking: a technical report

Authors

  • Andrew T. McKenzie Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Friedman Brain Institute, Departments of Pathology, Neuroscience, and Artificial Intelligence & Human Health, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Neuropathology Brain Bank & Research Core and Ronald M. Loeb Center for Alzheimer's Disease, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Department of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, New York, USA https://orcid.org/0000-0001-7462-4340
  • Emma Woodoff-Leith Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Friedman Brain Institute, Departments of Pathology, Neuroscience, and Artificial Intelligence & Human Health, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Neuropathology Brain Bank & Research Core and Ronald M. Loeb Center for Alzheimer's Disease, Icahn School of Medicine at Mount Sinai, New York, New York, USA
  • Diana Dangoor Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Friedman Brain Institute, Departments of Pathology, Neuroscience, and Artificial Intelligence & Human Health, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Neuropathology Brain Bank & Research Core and Ronald M. Loeb Center for Alzheimer's Disease, Icahn School of Medicine at Mount Sinai, New York, New York, USA
  • Alessandra Cervera Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Friedman Brain Institute, Departments of Pathology, Neuroscience, and Artificial Intelligence & Human Health, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Neuropathology Brain Bank & Research Core and Ronald M. Loeb Center for Alzheimer's Disease, Icahn School of Medicine at Mount Sinai, New York, New York, USA
  • Hadley Ressler Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Friedman Brain Institute, Departments of Pathology, Neuroscience, and Artificial Intelligence & Human Health, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Neuropathology Brain Bank & Research Core and Ronald M. Loeb Center for Alzheimer's Disease, Icahn School of Medicine at Mount Sinai, New York, New York, USA
  • Kristen Whitney Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Friedman Brain Institute, Departments of Pathology, Neuroscience, and Artificial Intelligence & Human Health, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Neuropathology Brain Bank & Research Core and Ronald M. Loeb Center for Alzheimer's Disease, Icahn School of Medicine at Mount Sinai, New York, New York, USA
  • Kristen Dams-O’Connor Department of Rehabilitation and Human Performance, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Department of Neurology, Icahn School of Medicine at Mount Sinai, New York, New York, USA
  • Zhuhao Wu Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Department of Cell, Developmental, and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Black Family Stem Cell Institute, Icahn School of Medicine at Mount Sinai, New York, New York, USA
  • Elizabeth M.C. Hillman Laboratory for Functional Optical Imaging, Departments of Biomedical Engineering and Radiology and the Mortimer B. Zuckerman Mind Brain Behavior Institute, Columbia University, New York, New York, USA
  • Alan C. Seifert Biomedical Engineering and Imaging Institute, Department of Diagnostic, Molecular and Interventional Ra-diology, and Graduate School of Biomedical Sciences, Icahn School of Medicine at Mount Sinai, New York, New York, USA
  • John F. Crary Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Friedman Brain Institute, Departments of Pathology, Neuroscience, and Artificial Intelligence & Human Health, Icahn School of Medicine at Mount Sinai, New York, New York, USA; Neuropathology Brain Bank & Research Core and Ronald M. Loeb Center for Alzheimer's Disease, Icahn School of Medicine at Mount Sinai, New York, New York, USA

DOI:

https://doi.org/10.17879/freeneuropathology-2022-4368

Keywords:

Brain banking, Perfusion fixation, Perfusion pressure, Postmortem interval, Tissue morphology, RNAscope, Ex vivo neuroimaging

Abstract

Perfusion fixation is a well-established technique in animal research to improve preservation quality in the study of many tissues, including the brain. There is a growing interest in using perfusion to fix postmortem human brain tissue to achieve the highest fidelity preservation for downstream high-resolution morphomolecular brain mapping studies. Numerous practical barriers arise when applying perfusion fixation in brain banking settings, including the large mass of the organ, degradation of vascular integrity and patency prior to the start of the procedure, and differing investigator goals sometimes necessitating part of the brain to be frozen. As a result, there is a critical need to establish a perfusion fixation procedure in brain banking that is flexible and scalable. This technical report describes our approach to developing an ex situ perfusion fixation protocol. We discuss the challenges encountered and lessons learned while implementing this procedure. Routine morphological staining and RNA in situ hybridization data show that the perfused brains have well-preserved tissue cytoarchitecture and intact biomolecular signal. However, it remains uncertain whether this procedure leads to improved histology quality compared to immersion fixation. Additionally, ex vivo magnetic resonance imaging (MRI) data suggest that the perfusion fixation protocol may introduce imaging artifacts in the form of air bubbles in the vasculature. We conclude with further research directions to investigate the use of perfusion fixation as a rigorous and reproducible alternative to immersion fixation for the preparation of postmortem human brains.

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Published

2022-09-28

How to Cite

McKenzie, A. T., Woodoff-Leith, E., Dangoor, D., Cervera, A., Ressler, H., Whitney, K., Dams-O’Connor, K., Wu, Z., Hillman, E. M., Seifert, A. C., & Crary, J. F. (2022). Ex situ perfusion fixation for brain banking: a technical report. Free Neuropathology, 3, 22. https://doi.org/10.17879/freeneuropathology-2022-4368
صندلی اداری سرور مجازی ایران Decentralized Exchange

Issue

Vol. 3 (2022)

Section

Original Papers

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Copyright (c) 2022 Andrew T. McKenzie, Emma Woodoff-Leith, Diana Dangoor, Alessandra Cervera, Hadley Ressler, Kristen Whitney, Kristen Dams-O’Connor, Zhuhao Wu, Elizabeth M.C. Hillman, Alan C. Seifert, John F. Crary

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This work is licensed under a Creative Commons Attribution 4.0 International License.

Papers are published open access under the Creative Commons BY 4.0 license. This license lets others distribute, remix, adapt, and build upon your work, even commercially, as long as they credit you for the original creation. Data included in the article are made available under the CC0 1.0 Public Domain Dedication waiver, unless otherwise stated, meaning that all copyrights are waived.

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