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Abstract. For efficient analysis and characterization of biological ice nuclei underimmersion freezing conditions, we developed the Twin-plate Ice Nucleation Assay(TINA) for high-throughputdroplet freezing experiments, in which the temperature profile and freezingof each droplet is tracked by an infrared detector. In the fully automatedsetup, a couple of independently cooled aluminum blocks carrying two 96-wellplates and two 384-well plates, respectively, are available to study icenucleation and freezing events simultaneously in hundreds of microliter-rangedroplets (0.1–40  µ L). A cooling system with two refrigerantcirculation loops is used for high-precision temperature control (uncertainty  K), enabling measurements over a wide range of temperatures( ∼  272–233 K) at variable cooling rates (up to 10 K min −1) . The TINA instrument was tested and characterized in experiments withbacterial and fungal ice nuclei (IN) from Pseudomonas syringae (Snomax®) and Mortierella alpina, exhibiting freezing curves in good agreement with literaturedata. Moreover, TINA was applied to investigate the influence of chemicalprocessing on the activity of biological IN, in particular the effects ofoxidation and nitration reactions. Upon exposure ofSnomax® to O3 and NO2 , the cumulativenumber of IN active at 270–266 K decreased by more than 1 order ofmagnitude. Furthermore, TINA was used to study aqueous extracts ofatmospheric aerosols, simultaneously investigating a multitude of samplesthat were pre-treated in different ways to distinguish different kinds ofIN. For example, heat treatment and filtration indicated that mostbiological IN were larger than 5  µ m. The results confirm that TINA issuitable for high-throughput experiments and efficient analysis ofbiological IN in laboratory and field samples.

atmospheric measurement techniques

Twin-plate Ice Nucleation Assay (TINA) with infrared detection for high-throughput droplet freezing experiments with biological ice nuclei in laboratory and field samples