We firstly demonstrate the three-dimensional (3D) measurement of a nanometer-sized sphere held in optical tweezers in water using an in-line digital holographic microscope with a green light emitting diode. Suppressing the movement with optical tweezers enabled us to detect the three-dimensional position of a polystyrene sphere with a diameter of $200\mathrm{nm}$ . The positioning resolutions of the microscope were $3.2\mathrm{nm}$ in the transverse direction and $3.4\mathrm{nm}$ in the axial direction, from the standard deviation of measurements of the $200\mathrm{nm}$ sphere fixed on glass. Changes in the Brownian motion in response to a change in the trapping laser power were measured. We also demonstrated that this holographic measurement is an effective method for determining the threshold power of the optical trapping.

© 2011 Optical Society of America

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