Circa 2015, brain immortality via aldehyde-stabilized cryopreservation.
Here we describe a new cryobiological technique for neurobiological research, called aldehyde stabilized cryopreservation. This technique demonstrates the importance and utility of advanced cryopreservation technology to the neurobiological community. ASC is an innovative brain-banking method that combines ice-free storage for long periods of time with high-resolution images. We perfused glutaraldehyde fixated rabbit and pigs brains for several hours to demonstrate the feasibility ASC. Then we slowly perfused increasing concentrations ethylene glycol in a similar manner as techniques used for cryopreservation of whole organs. We vitrified the brains once a concentration of 65% w/v was reached. This will allow them to be stored for an indefinite period at -135 degrees Celsius. The vitrified brains were then rewarmed, and the cryoprotectant was removed by either perfusion or diffusion. ASC-processed cerebral tissue was evaluated by scanning electron microscopy and FIB-SEM imaging. The preservation was excellent in both species: the processes were traceable, and the synapses were sharp. Aldehyde stabilized cryopreservation offers many advantages to other brain-banking methods: chemicals can be delivered via perfusion which allows for easy scaling of brains of all sizes; vitrification guarantees that the ultrastructure will not degrade over long storage periods; and the cryoprotectant is easily removed, resulting in a perfusable, aldehyde preserved brain suitable for various brain assays.