Fig. 5.

Freezing temperatures of purified water droplet ensembles at a range of volumes with an empirical trend. Boxes denote 25th – 75th percentiles, bars denote medians and whiskers denote range of values. We have plotted literature data [4,5,13,25,34,37] of ice nucleation temperatures of pure water aliquots in various containers and spanning volumes from sub-pL to tens of mL along with a homogenous freezing parameterisation [16]. We also show new data for 100 μL droplets in a 96-well plate (from Fig. 2) and data for 1.0 mL in cryovials (Corning 2.0 mL capacity, Cat No. 430488) and 70 mL in cryobags (Milenyi CD250). We used Hyclone cell culture water. The cryovials assay was performed using the IR-NIPI as previously described (n = 31); while for the cryobags the ice nucleation temperatures (n = 6) were determined using a thermocouple as the bags were cooled while inside aluminium cassettes.