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Aldehyde-stabilized cryopreservation
Open accessAbstract
We describe here a new cryobiological and neurobiological technique, aldehyde-stabilized cryopreservation (ASC), which demonstrates the relevance and utility of advanced cryopreservation science for the neurobiological research community. ASC is a new brain-banking technique designed to facilitate neuroanatomic research such as connectomics research, and has the unique ability to combine stable long term ice-free sample storage with excellent anatomical resolution. To demonstrate the feasibility of ASC, we perfuse-fixed rabbit and pig brains with a glutaraldehyde-based fixative, then slowly perfused increasing concentrations of ethylene glycol over several hours in a manner similar to techniques used for whole organ cryopreservation. Once 65% w/v ethylene glycol was reached, we vitrified brains at -135 °C for indefinite long-term storage. Vitrified brains were rewarmed and the cryoprotectant removed either by perfusion or gradual diffusion from brain slices. We evaluated ASC-processed brains by electron microscopy of multiple regions across the whole brain and by Focused Ion Beam Milling and Scanning Electron Microscopy (FIB-SEM) imaging of selected brain volumes. Preservation was uniformly excellent: processes were easily traceable and synapses were crisp in both species. Aldehyde-stabilized cryopreservation has many advantages over other brain-banking techniques: chemicals are delivered via perfusion, which enables easy scaling to brains of any size; vitrification ensures that the ultrastructure of the brain will not degrade even over very long storage times; and the cryoprotectant can be removed, yielding a perfusable aldehyde-preserved brain which is suitable for a wide variety of brain assays.
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Cites
Structural and molecular interrogation of intact biological systems
Chung, K., Wallace, J., Kim, S.-Y., Kalyanasundaram, S., Andalman, A. S., Davidson, T. J., Mirzabekov, J. J., Zalocusky, K. A., Mattis, J., Denisin, A. K., et al. (2013) · Nature · doi:10.1038/nature12107 · Cited by 2,008
The essential role of hippocampal CA1 NMDA receptor-dependent synaptic plasticity in spatial memory.
Tsien, J. Z., Huerta, P. T., & Tonegawa, S. (1996) · Cell · doi:10.1016/s0092-8674(00)81827-9 · Cited by 1,782
Chen, F., Tillberg, P. W., & Boyden, E. S. (2015) · Science · doi:10.1126/science.1260088 · Cited by 1,516
(2006) · doi:10.1093/acprof:oso/9780195100273.001.0001 · Cited by 1,186
Vitrification as an approach to cryopreservation
Fahy, G. M., MacFarlane, D. R., Angell, C. A., & Meryman, H. T. (1984) · Cryobiology · doi:10.1016/0011-2240(84)90079-8 · Cited by 983
Self-citationContributions and challenges for network models in cognitive neuroscience
Sporns, O. (2014) · Nature Neuroscience · doi:10.1038/nn.3690 · Cited by 767
Fine Structure Immunocytochemistry
Griffiths, G. W. (1993) · Springer eBooks · doi:10.1007/978-3-642-77095-1 · Cited by 699
Fixation for Electron Microscopy
(1981) · Elsevier eBooks · doi:10.1016/b978-0-12-333920-1.x5001-0 · Cited by 600
FIXATION OF NEURAL TISSUES FOR ELECTRON MICROSCOPY BY PERFUSION WITH SOLUTIONS OF OSMIUM TETROXIDE
Palay, S. L., McGee-Russell, S. M., Gordon, S., & Grillo, M. A. (1962) · The Journal of Cell Biology · doi:10.1083/jcb.12.2.385 · Cited by 534
A technicolour approach to the connectome
Lichtman, J. W., Livet, J., & Sanes, J. R. (2008) · Nature reviews. Neuroscience · doi:10.1038/nrn2391 · Cited by 328
Flagged first, then by citations
Research works printed but not linked: 2
- Fahy, 1984, Histological cryoprotection of rat and rabbit brains, CryoLetters, 5, 33
- Fahy, 1980, Activation of alpha adrenergic vasoconstrictor response in kidneys stored at −30 °C for up to 8 days, CryoLetters, 1, 312
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Berger, D. R., Seung, H. S., & Lichtman, J. W. (2018) · Frontiers in Neural Circuits · doi:10.3389/fncir.2018.00088 · Cited by 206
Perfusion fixation in brain banking: a systematic review
McFadden, W. C., Ressler, H. W., Richter, F., Soudant, C., Bryce, C., Hof, P. R., Fowkes, M. E., Crary, J. F., & McKenzie, A. T. (2019) · Acta Neuropathologica Communications · doi:10.1186/s40478-019-0799-y · Cited by 94
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Carneiro, I. C., Carvalho, S. D., Henrique, R., Oliveira, L. M. C., & Tuchin, V. V. (2018) · Journal of Biophotonics · doi:10.1002/jbio.201800333 · Cited by 33
Yamada, H., Yamaguchi, M., Shimizu, K., Murayama, S. Y., Mitarai, S., Sasakawa, C., & Chibana, H. (2017) · Microscopy · doi:10.1093/jmicro/dfx019 · Cited by 26
McKenzie, A. T. (2019) · OSF Preprints (OSF Preprints) · doi:10.31219/osf.io/8zd4e · Cited by 25
In situ X-ray-assisted electron microscopy staining for large biological samples
Ströh, S., Hammerschmith, E. W., Tank, D. W., Seung, H. S., & Wanner, A. A. (2022) · eLife · doi:10.7554/elife.72147 · Cited by 23
Structural brain preservation: a potential bridge to future medical technologies
McKenzie, A. T., Zeleznikow-Johnston, A., Sparks, J. S., Nnadi, O., Smart, J., Wiley, K., Cerullo, M. A., de Wolf, A., Minerva, F., Risco, R., et al. (2024) · Frontiers in Medical Technology · doi:10.3389/fmedt.2024.1400615 · Cited by 15
de Wolf, A., Phaedra, C., Perry, R. M., & Maire, M. (2019) · Rejuvenation Research · doi:10.1089/rej.2019.2225 · Cited by 12
Dein, S. L. (2021) · Journal of Religion and Health · doi:10.1007/s10943-020-01166-6 · Cited by 11
Functional recovery of the adult murine hippocampus after cryopreservation by vitrification
German, A., Akdaş, E. Y., Flügel‐Koch, C., Erterek, E., Frischknecht, R., Fejtová, A., Winkler, J., Alzheimer, C., & Zheng, F. (2026) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2516848123 · Cited by 10
Principles of Vitrification as a Method of Cryopreservation in Reproductive Biology and Medicine
Fahy, G. M. (2021) · Cambridge University Press eBooks · doi:10.1017/9781108784368.007 · Cited by 8
Self-citationMarin, F. (2017) · arXiv (Cornell University) · doi:10.48550/arxiv.1708.08649 · Cited by 8
Biostasis: A Roadmap for Research in Preservation and Potential Revival of Humans
McKenzie, A. T., Wowk, B. G., Arkhipov, A. S., Wróbel, B., Cheng, N., & Kendziorra, E. F. (2024) · Brain Sciences · doi:10.3390/brainsci14090942 · Cited by 7
The Ethics of Exponential Life Extension through Brain Preservation
Cerullo, M. A. (2016) · Journal of Ethics and Emerging Technologies · doi:10.55613/jeet.v26i1.54 · Cited by 8
Ekpo, M. D., Boafo, G. F., Gambo, S. S., Hu, Y., Liu, X., Xie, J., & Tan, S. (2022) · Frontiers in Veterinary Science · doi:10.3389/fvets.2022.877163 · Cited by 6
Cryopreservation - Applications and Challenges
Quain, M. (2022) · Biomedical engineering · doi:10.5772/intechopen.100666 · Cited by 6
Warner, R. M., Yang, J., Drake, A., Lee, Y. J., Nemanic, S., Scott, D., & Higgins, A. Z. (2023) · PeerJ · doi:10.7717/peerj.16323 · Cited by 4
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Ultrastructural and Histological Cryopreservation of Mammalian Brains by VitrificationPreprint
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German, A., & Tretter, M. (2025) · Neuroethics · doi:10.1007/s12152-025-09584-7 · Cited by 3
McBride, J. (2022) · Journal of Posthumanism · doi:10.33182/joph.v2i1.1788 · Cited by 3
Overcoming Perfusion Impairment in the Ischemic Brain: A Review
McKenzie, A. T., de Wolf, A., Grotemeyer, A., Kendziorra, E., & German, A. (2026) · NeuroSci · doi:10.3390/neurosci7040089 · Cited by 2
Future Projections and Fallouts of Exponential Longevity and Revival from Cryopreservation
Nikhra, V. (2018) · Research in Medical & Engineering Sciences · doi:10.31031/rmes.2018.06.000641 · Cited by 2
James, R. I., Verma, R., Johnson, L. R., Abi Manesh, S., Jayakumar, J., Sen, M., Joseph, J., Kumarasami, R., Mitra, P. P., Sivaprakasam, M., et al. (2023) · American Journal of Forensic Medicine & Pathology · doi:10.1097/paf.0000000000000871 · Cited by 2
Physician estimates of the feasibility of preserving the dying for future revival
Zeleznikow-Johnston, A., Kendziorra, E. F., & McKenzie, A. T. (2026) · PLoS ONE · doi:10.1371/journal.pone.0348216 · Cited by 1
Cryopreservation of aldehyde-fixed whole brains
Garrood, M., Keberle, A., Slaughter, A., Sowa, A., Thorn, E. L., Sanctis, C. D., Farrell, K. W., Crary, J. F., & McKenzie, A. T. (2026) · PLoS ONE · doi:10.1371/journal.pone.0344932 · Cited by 1
Slaughter, A., Garrood, M., Hedden, K., Sowa, A., Janssen, W., Thorn, E. L., Sanctis, C. D., Farrell, K., Crary, J. F., & McKenzie, A. T. (2026) · Ultrastructural Pathology · doi:10.1080/01913123.2026.2745999 · Cited by 1
Assessment of ethical aspects of cryonics: An emerging technology
Shanbhag, V. K. L. (2017) · Journal of the Scientific Society · doi:10.4103/jss.jss_19_17 · Cited by 1
Song, A., LaVergne, A., & Wróbel, B. (2026) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.64898/2026.03.04.709724 · Cited by 1
Self-citationThe Paradoxical Death through Cryothanasia or Playing God. A Moral-Theological Approach
Vulcănescu, S.-G. (2022) · Journal of Intercultural Management and Ethics · doi:10.35478/jime.2022.1.04
Purism: Desire as the Ultimate Value, Part Two An Appeal to Intuition
Primus (2023) · Philosophical Papers and Review · doi:10.5897/ppr2022.0222
DIY peristaltic perfusion pump and rotarod apparatuses from common and recycled materials
Almuslehi, M. S. M., & Coorssen, J. R. (2024) · Instrumentation Science & Technology · doi:10.1080/10739149.2024.2344713
Practitioner forecasts of technological progress in biostasisPreprint
McKenzie, A. T., Cerullo, M. A., Farahani, N., Sparks, J. S., Londoño, T., de Wolf, A., Dziennis, S., Wróbel, B., German, A., Kendziorra, E. F., et al. (2025) · arXiv (Cornell University) · doi:10.48550/arxiv.2507.17274
Hait, A., & Balaji, S. (2026) · Research Square · doi:10.21203/rs.3.rs-8284559/v1
Reanimación cerebral y vida después de la muerte: una revisión narrativa
Guzmán-Ríos, E. D., Guzman, K., Madera, E., R., I. J. G., M., R. R. Z., Olvera-Castro, J. F., Flores-Reyes, M. F., Moroscoso, R., & Guzmán, V. D. (2026) · ALERTA Revista Científica del Instituto Nacional de Salud · doi:10.5377/alerta.v9i3.23306
German, A., Grotemeyer, A., Koller, K., Eckstein, M., Wolff, H., Rampp, S., Süß, P., Müller, V. C., Birnbacher, D., Winkler, J., et al. (2026) · BMC Medical Ethics · doi:10.1186/s12910-026-01586-1
Mass Transfer in Cryopreservation at Multiple Scales: Cells, Tissues, and Organs
Benson, J. D., Consiglio, A. N., Powell‐Palm, M. J., Thompson, R. M., Giers, M. B., & Higgins, A. Z. (2026) · Methods in molecular biology · doi:10.1007/978-1-0716-5416-3_3
Ultrastructural comparison of fixation and cryopreservation methods for brain preservationPreprint
German, A., Flügel‐Koch, C., Paulsen, F., Winkler, J., & McKenzie, A. T. (2026) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.64898/2026.09.18.751896
Metadata
- Authors
- Song, A., & Fahy, G. M.
- Affiliations
- 21st Century Medicine (United States) (US)
- Corresponding author
- Song, Aurelia
- Venue
- Cryobiology
- Publisher
- Elsevier BV
- Published
- 2015-12
- Volume
- 71
- Issue
- 3
- Pages
- 448-458
- ISSN
- 0011-2240 (ISSN-L)
- OpenAlex
- W1770441180
- Cited by
- 42
- References
- 37
- Citations by year
- 2026: 12 · 2025: 3 · 2024: 3 · 2023: 3 · 2022: 6 · 2021: 2 · 2019: 4 · 2018: 4 · 2017: 4 · 2016: 1
- FWCI
- 2.89
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Cryobiology (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Brain Preservation Foundation
- Topics
- Health Sciences › Medicine › Surgery › Organ Transplantation Techniques and Outcomes
- Health Sciences › Medicine › Reproductive Medicine › Reproductive Biology and Fertility
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
- Keywords
- brain banking, glutaraldehyde fixation, cryoprotectant removal, ultrastructure preservation, connectomics, perfusion fixation, rabbit brain, pig brain
- MeSH terms
- Animals, Brain, Cryoprotective Agents, Glutaral, Rabbits, Swine, Tissue Banks, Cryopreservation, Ethylene Glycol, Vitrification
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