Calibration of optical tweezers with non-spherical probes via high-resolution detection of Brownian motion

Optical tweezers are commonly used and powerful tools to perform force measurements on the piconewton scale and to detect nanometer-scaled displacements. However, the precision of these instruments relies to a great extent on the accuracy of the calibration method. A well-known calibration procedure is to record the stochastic motion of the trapped particle and compare its statistical behavior with the theory of the Brownian motion in a harmonic potential. Here we present an interactive calibration software which allows for the simultaneous fitting of three different statistical observables (power spectral density, mean square displacement and velocity autocorrelation function) calculated from the trajectory of the probe to enhance fitting accuracy. The fitted theory involves the hydrodynamic interactions experimentally observable at high sampling rates. Furthermore, a qualitative extension is included in our model to handle the thermal fluctuations in the orientation of optically trapped asymmetric objects. The presented calibration methodology requires no prior knowledge of the bead size and can be applied to non-spherical probes as well. The software was validated on synthetic and experimental data. Program summary Program title: PFMCal Catalogue identifier: AEXH_v1_0 Program summary URL: http://cpc.cs.qub.ac.ukisummaries/AEXH_v1_0.html Program obtainable from: CPC Program Library, Queen's University, Belfast, N. Ireland Licensing provisions: Standard CPC licence, http://cpc.cs.qub.ac.uk/licence/licence.html No. of lines in distributed program, including test data, etc.: 206,399 No. of bytes in distributed program, including test data, etc.: 10,319,465 Distribution format: tar.gz Programming language: MatLab 2011a (Math Works Inc.). Computer: General computer running MatLab (MathWorks Inc.), using Statistics Toolbox. Operating system: Any which supports Matlab using Statistics Toolbox. RAM: 10 MB Classification: 3, 4.9, 18, 23. Nature of problem: Calibration of optical tweezers by measuring the Brownian motion of the trapped object. The voltage-to-displacement ratio of the detection system (the inverse of the sensitivity), the stiffness of the trap and the size of the bead are obtained via the simultaneous fitting of the power spectral density (PSD), mean square displacement (MSD) and velocity autocorrelation (VAF) functions calculated from the trajectory. The calibration can be performed for non-spherical probes as well. Solution method: Initialization points for all parameters are inferred from characteristic features of the statistical observables (PSD, MSD and VAF) based on the method developed by Grimm et al. in [1]. Theoretical functions for the PSD, the MSD and the VAF are calculated from the model of Brownian motion confined by a harmonic potential taking hydrodynamic interactions into consideration [2-4]. This calibration methodology has been successfully used in actual experiments with micro-spheres [5, 6]. Calculated functions are f