We explain and demonstrate a new method of force and position calibrations for optical tweezers with back-focal-plane photodetection. The method combines power spectral measurements of thermal motion and the response to a sinusoidal motion of a translation stage. It consequently does not use the drag coefficient of the trapped object as an input. Thus, neither the viscosity, nor the size of the trapped object, nor its distance to nearby surfaces needs to be known. The method requires only a low level of instrumentation and can be applied in situ in all spatial dimensions. It is both accurate and precise: true values are returned, with small error bars. We tested this experimentally, near and far from surfaces in the lateral directions. Both position and force calibrations were accurate to within 3%. To calibrate, we moved the sample with a piezoelectric translation stage, but the laser beam could be moved instead, e.g., by acousto-optic deflectors. Near surfaces, this precision requires an improved formula for the hydrodynamical interaction between an infinite plane and a microsphere in nonconstant motion parallel to it. We give such a formula.
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October 2006
Research Article|
October 09 2006
Calibration of optical tweezers with positional detection in the back focal plane
Simon F. Tolić-Nørrelykke;
Simon F. Tolić-Nørrelykke
Max Planck Institute for the Physics of Complex Systems
, Nöthnitzer Strasse 38, 01187 Dresden, Germany and European Laboratory for Non-linear Spectroscopy
, via Nello Carrara 1, 50019 Sesto Fiorentino, Florence, Italy
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Erik Schäffer;
Max Planck Institute of Molecular Cell Biology and Genetics
, Pfotenhauerstrasse 108, 01307 Dresden, Germany
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Jonathon Howard;
Jonathon Howard
Max Planck Institute of Molecular Cell Biology and Genetics
, Pfotenhauerstrasse 108, 01307 Dresden, Germany
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Francesco S. Pavone;
Francesco S. Pavone
European Laboratory for Non-linear Spectroscopy
, via Nello Carrara 1, 50019 Sesto Fiorentino, Florence, Italy
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Frank Jülicher;
Frank Jülicher
Max Planck Institute for the Physics of Complex Systems
, Nöthnitzer Strasse 38, 01187 Dresden, Germany
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Henrik Flyvbjerg
Henrik Flyvbjerg
Isaac Newton Institute for Mathematical Sciences
, Cambridge CB3 0EH United Kingdom; Biosystems Department, Risø National Laboratory
, DK-4000 Roskilde, Denmark; and Danish Polymer Centre, Risø National Laboratory
, DK-4000 Roskilde, Denmark
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Rev. Sci. Instrum. 77, 103101 (2006)
Article history
Received:
February 13 2006
Accepted:
August 26 2006
Citation
Simon F. Tolić-Nørrelykke, Erik Schäffer, Jonathon Howard, Francesco S. Pavone, Frank Jülicher, Henrik Flyvbjerg; Calibration of optical tweezers with positional detection in the back focal plane. Rev. Sci. Instrum. 1 October 2006; 77 (10): 103101. https://doi.org/10.1063/1.2356852
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