The DOI matches its Crossref record; 39 of the 39 references it cites were found in a registry or an index.
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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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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