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Atmospheric Measurement Techniques An interactive open-access journal of the European Geosciences Union
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Discussion papers | Copyright
https://doi.org/10.5194/amt-2018-230
© Author(s) 2018. This work is distributed under
the Creative Commons Attribution 4.0 License.

Research article 24 Jul 2018

Research article | 24 Jul 2018

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This discussion paper is a preprint. It is a manuscript under review for the journal Atmospheric Measurement Techniques (AMT).

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

Anna T. Kunert1, Mark Lamneck2, Frank Helleis2, Mira L. Pöhlker1, Ulrich Pöschl1, and Janine Fröhlich-Nowoisky1 Anna T. Kunert et al.
  • 1Multiphase Chemistry Department, Max Planck Institute for Chemistry, 55128 Mainz, Germany
  • 2Instrument Development and Electronics, Max Planck Institute for Chemistry, 55128 Mainz, Germany

Abstract. For efficient analysis and characterization of biological ice nuclei under immersion freezing conditions, we developed a Twin-plate Ice Nucleation Assay (TINA) for high-throughput droplet freezing experiments, in which the temperature gradient and freezing of each droplet is tracked by an infrared detector. In the fully automated setup, a couple of independently cooled aluminum blocks carrying two 96-well plates and two 384-well plates, respectively, are available to study ice nucleation and freezing events simultaneously in hundreds of microliter range droplets (0.1–40µL). A cooling system with two refrigerant circulation loops is used for high-precision temperature control (deviations <0.5K), enabling measurements over a wide range of temperatures (~233–270K) at variable cooling rates (up to 10Kmin−1 and more).

The TINA instrument was tested and characterized in experiments with bacterial and fungal ice nuclei (IN) from Pseudomonas syringae (Snomax®) and Mortierella alpina, exhibiting freezing curves in good agreement with literature data. Moreover, TINA was applied to investigate the influence of chemical processing on the activity of biological IN, in particular the effects of oxidation and nitration reactions. Upon exposure of Snomax® samples to O3 and NO2, the concentration of IN active around 269K (−4°C, “class A”) decreased by about two orders of magnitude, while the concentration of IN active around 265K (−8°C, “class C”) decreased by about one order of magnitude. Furthermore, TINA was used to study aqueous extracts of atmospheric aerosols, simultaneously investigating a multitude of samples that were treated in different ways to distinguish different kinds of IN. For example, heat treatment and filtration experiments indicated that highly efficient biological IN were smaller than 0.1µm, and many IN active between 263K (−10°C) and 256K (−14°C) were heat-resistant and larger than 5µm. The results confirm that TINA is suitable for high-throughput experiments and efficient analysis of biological IN in laboratory and field samples.

Anna T. Kunert et al.
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Status: final response (author comments only)
Status: final response (author comments only)
AC: Author comment | RC: Referee comment | SC: Short comment | EC: Editor comment
Anna T. Kunert et al.
Anna T. Kunert et al.
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A new twin-plate ice nucleation assay with infrared detection for high-throughput droplet freezing experiments in microliter-sized droplets is introduced, which was tested and characterized with bacterial and fungal IN. It was applied to investigate the influence of chemical processing on the activity of biological IN, and aqueous extracts of atmospheric aerosols were studied for IN activity.
A new twin-plate ice nucleation assay with infrared detection for high-throughput droplet...
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