Figure 3. Size exclusion chromatography combined with an ice nucleation assay to study the size of INPs in the fungus Fusarium avenaceum.

(Left) Experimental schematic of the fractionation procedure. Initially, 96 × 300 μL fractions are collected from the Superose 6 10/300 GL column. A droplet freezing experiment is then performed using a 1 μL droplet from each of the fractions. (Right) UV absorbance (black trace) and freezing temperature of eluted fractions (blue trace) after passing through a Superose 6 column. The appearance of elevated nucleation temperatures can be seen concurrent to elevated absorbance in the eluate at flow through volumes of 6–11 mls (~1000–5000+ kDa).