Camera-based three-dimensional real-time particle tracking at kHz rates and Ångström accuracy

Alexander Huhle, Daniel Klaue, Hergen Brutzer, Peter Daldrop, Sihwa Joo, Oliver Otto, Ulrich F. Keyser and Ralf Seidel ()
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Alexander Huhle: Biotechnology Center, Technische Universität Dresden
Daniel Klaue: Biotechnology Center, Technische Universität Dresden
Hergen Brutzer: Biotechnology Center, Technische Universität Dresden
Peter Daldrop: Institute for Molecular Cell Biology, Westfälische Wilhelms-Universität Münster
Sihwa Joo: Biotechnology Center, Technische Universität Dresden
Oliver Otto: University of Cambridge, Cavendish Laboratory, JJ Thomson Avenue, Cambridge CB3 0HE, UK
Ulrich F. Keyser: University of Cambridge, Cavendish Laboratory, JJ Thomson Avenue, Cambridge CB3 0HE, UK
Ralf Seidel: Biotechnology Center, Technische Universität Dresden

Nature Communications, 2015, vol. 6, issue 1, 1-8

Abstract: Abstract Optical and magnetic tweezers are widely employed to probe the mechanics and activity of individual biomolecular complexes. They rely on micrometre-sized particles to detect molecular conformational changes from the particle position. Real-time particle tracking with Ångström accuracy has so far been only achieved using laser detection through photodiodes. Here we demonstrate that camera-based imaging can provide a similar performance for all three dimensions. Particle imaging at kHz rates is combined, with real-time data processing being accelerated by a graphics-processing unit. For particles that are fixed in the sample cell we can detect 3-Å-sized steps that are introduced by cell translations at rates of 10 Hz, while for DNA-tethered particles 5 Å steps at 1 Hz can be resolved. Moreover, 20 particles can be tracked in parallel with comparable accuracy. Our approach provides a simple and robust way for high-resolution tweezer experiments using multiple particles at a time.

Date: 2015
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DOI: 10.1038/ncomms6885

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