Optimizing bead size reduces errors in force measurements in optical traps.

Optical traps are used to measure force (F) over a wide range (0.01 to 1,000 pN). Variations in bead radius (r) hinder force precision since trap stiffness (k(trap)) varies as r3 when r is small. Prior work has shown k(trap) is maximized when r is approximately equal to the beam waist (w0), which on our instrument was ~400 nm when trapping with a 1064-nm laser. In this work, we show that by choosing r ≈w0, we improved the force precision by 2.8-fold as compared to a smaller bead (250 nm). This improvement in force precision was verified by pulling on a canonical DNA hairpin. Thus, by using an optimum bead size, one can simultaneously maximize k(trap) while minimizing errors in F.

[1]  Michelle D. Wang,et al.  Stretching DNA with optical tweezers. , 1997, Biophysical journal.

[2]  M. Rief,et al.  The Complex Folding Network of Single Calmodulin Molecules , 2011, Science.

[3]  Christoph F. Schmidt,et al.  Calibrating bead displacements in optical tweezers using acousto-optic deflectors , 2006 .

[4]  Alexander Rohrbach,et al.  Stiffness of optical traps: quantitative agreement between experiment and electromagnetic theory. , 2005, Physical review letters.

[5]  M. Woodside,et al.  Direct observation of multiple misfolding pathways in a single prion protein molecule , 2012, Proceedings of the National Academy of Sciences.

[6]  Ashley R. Carter,et al.  Precision surface-coupled optical-trapping assay with one-basepair resolution. , 2009, Biophysical journal.

[7]  Ashley R. Carter,et al.  Stabilization of an Optical Microscope to 0.1 Nm in Three Dimensions , 2022 .

[8]  Carlos Bustamante,et al.  Optical-trap force transducer that operates by direct measurement of light momentum. , 2003, Methods in enzymology.

[9]  Francesco S. Pavone,et al.  Calibration of optical tweezers with positional detection in the back focal plane , 2006, physics/0603037.

[10]  J. Liphardt,et al.  Reversible Unfolding of Single RNA Molecules by Mechanical Force , 2001, Science.

[11]  S. Chu,et al.  Quantitative measurements of force and displacement using an optical trap. , 1996, Biophysical journal.

[12]  Derek N. Fuller,et al.  DNA as a metrology standard for length and force measurements with optical tweezers. , 2006, Biophysical journal.

[13]  Christoph F. Schmidt,et al.  Direct observation of kinesin stepping by optical trapping interferometry , 1993, Nature.

[14]  D. Herschlag,et al.  Nanomechanical measurements of the sequence-dependent folding landscapes of single nucleic acid hairpins. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[15]  T. Perkins,et al.  Measuring 0.1-nm motion in 1 ms in an optical microscope with differential back-focal-plane detection. , 2004, Optics letters.

[16]  Mario Montes-Usategui,et al.  Optimized back-focal-plane interferometry directly measures forces of optically trapped particles. , 2012, Optics express.

[17]  D. Grier A revolution in optical manipulation , 2003, Nature.

[18]  D. Herschlag,et al.  Direct Measurement of the Full, Sequence-Dependent Folding Landscape of a Nucleic Acid , 2006, Science.

[19]  Carlos Bustamante,et al.  Direct Observation of the Three-State Folding of a Single Protein Molecule , 2005, Science.

[20]  S. Block,et al.  Construction of multiple-beam optical traps with nanometer-resolution position sensing , 1996 .

[21]  Mario Montes-Usategui,et al.  A force detection technique for single-beam optical traps based on direct measurement of light momentum changes. , 2010, Optics express.

[22]  Yann R Chemla,et al.  Characterization of photoactivated singlet oxygen damage in single-molecule optical trap experiments. , 2009, Biophysical journal.

[23]  Mark J. Schnitzer,et al.  Single kinesin molecules studied with a molecular force clamp , 1999, Nature.

[24]  W. Greenleaf,et al.  High-resolution, single-molecule measurements of biomolecular motion. , 2007, Annual review of biophysics and biomolecular structure.

[25]  P. Nelson,et al.  Elasticity of short DNA molecules: theory and experiment for contour lengths of 0.6-7 microm. , 2007, Biophysical journal.

[26]  I. Tinoco,et al.  Equilibrium Information from Nonequilibrium Measurements in an Experimental Test of Jarzynski's Equality , 2002, Science.

[27]  Thomas T Perkins,et al.  Overstretching DNA at 65 pN does not require peeling from free ends or nicks. , 2011, Journal of the American Chemical Society.

[28]  C. Schmidt,et al.  Interference model for back-focal-plane displacement detection in optical tweezers. , 1998, Optics letters.

[29]  K. Svoboda,et al.  Biological applications of optical forces. , 1994, Annual review of biophysics and biomolecular structure.

[30]  Watt W. Webb,et al.  Measurement of small forces using an optical trap , 1994 .

[31]  Carlos Bustamante,et al.  Recent advances in optical tweezers. , 2008, Annual review of biochemistry.

[32]  M. Woodside,et al.  Single-Molecule Manipulation Using Optical Traps , 2009 .

[34]  G. I. Bell Models for the specific adhesion of cells to cells. , 1978, Science.

[35]  K. Neuman,et al.  Optical trapping. , 2004, The Review of scientific instruments.

[36]  E. Stelzer,et al.  Three‐dimensional high‐resolution particle tracking for optical tweezers by forward scattered light , 1999, Microscopy research and technique.

[37]  Colin Echeverría Aitken,et al.  An oxygen scavenging system for improvement of dye stability in single-molecule fluorescence experiments. , 2008, Biophysical journal.

[38]  Jonathon Howard,et al.  Optical trapping of coated microspheres. , 2008, Optics express.

[39]  Thomas T. Perkins,et al.  Optical traps for single molecule biophysics: a primer , 2009 .

[40]  A. Buosciolo,et al.  New calibration method for position detector for simultaneous measurements of force constants and local viscosity in optical tweezers , 2004 .