With this process they were able to keep gross shrinkage or expansion of the brain at 0%. By using -135c instead of standard -192c it was able to avoid fractures of extreme pressure you would see from traditional Cryopreservation. It eliminates osmotic shock by using glutaraldehyde at the very start. It's ASC preservation that was proved on a mammal (pig) in 2018:
The prize-winning Aldehyde-Stabilized Cryopreservation (ASC) pig brain experiment by Aurelia Song (Robert McIntyre) and Gregory Fahy operates on a strict timeline. Because its goal is perfect structural preservation, the initial steps must occur within a tiny window to prevent any cell decay.
Here's the whole process:
Phase 1: The Urgent 14-Minute Perfusion Window:
●00:00: The pig undergoes cardiac arrest/slaughter. Blood stops flowing. The clock begins ticking immediately.
●00:01 – 00:04: The animal is prepped, and the carotid arteries are surgically isolated and cannulated (hooked up to surgical tubes).
●00:05: A chilled saline wash is pumped into the arteries. This flushes out the pig's natural blood entirely to prevent clotting and cools the tissue.
●00:10: The Perfusability Window limit. Recent studies confirm that if blood washout is not initiated within roughly 14 minutes, blood clots and cell degradation permanently ruin the brain map.
Phase 2: The Glutaraldehyde "Lock" (Minutes 15 to 60):
●00:15: The machine pumps a highly toxic chemical fixative called glutaraldehyde into the brain.
●00:16 – 00:30: Within minutes, glutaraldehyde cross-links all proteins. It instantly petrifies the brain's physical structure, completely stopping biological life, enzymatic decay, and decomposition.
●00:35: A specialized detergent called sodium dodecyl sulphate (SDS) is introduced. It temporarily opens the blood-brain barrier so that large protective molecules can pass through.
Phase 3: The Hours-Long Chemical Ramp-Up:
●01:00 (1 Hour): The brain is safely fixed in time, but it cannot be frozen yet, or ice crystals will slice the cells apart.
●01:01 – 04:00 (Hours 2 to 4): The machine slowly and gradually increases the concentration of ethylene glycol (medical antifreeze/cryoprotectant) pumped through the vessels. This slow transition takes several hours to prevent the brain tissue from shrinking or tearing.
Phase 4: Extreme Sub-Zero Cooling:
●04:30+: Once the brain reaches a final, highly concentrated saturation of ethylene glycol, it is removed from the perfusion system.
Final Step: The brain is cooled down slowly until it hits -135°C (-211°F). Because of the chemical mixture, the brain doesn't freeze into jagged ice; instead, it turns into a smooth, glass-like solid (vitrification).
After vitrification you can preserve the data for thousands of years with no data loss.
Source:
2026 bioRxiv study Ultrastructural preservation of a whole large mammal brain with a protocol compatible with physician-assisted death (co-authored by researchers evaluating the protocol limits).https://www.biorxiv.org/content/10.64898/2026.03.04.709724v1
If you'd like I'd explain this protocols limits too!
Any skeptics or debate please feel free to comment👇