  Stable aqueous supercooling has shown significant potential as a technique
for human tissue preservation, food cold storage, conservation biology, and
beyond, but its stochastic nature has made its translation outside the
laboratory difficult. In this work, we present an isochoric nucleation
detection (INDe) platform for automated, high-throughput characterization of
aqueous supercooling at >1 mL volumes, which enables statistically-powerful
determination of the temperatures and time periods for which supercooling in a
given aqueous system will remain stable. We employ the INDe to investigate the
effects of thermodynamic, surface, and chemical parameters on aqueous
supercooling, and demonstrate that various simple system modifications can
significantly enhance supercooling stability, including isochoric
(constant-volume) confinement, hydrophobic container walls, and the addition of
even mild concentrations of solute. Finally, in order to enable informed design
of stable supercooled biopreservation protocols, we apply a statistical model
to estimate stable supercooling durations as a function of temperature and
solution chemistry, producing first-in-field supercooling stability maps for
four common cryoprotective solutes.
