We present a new technique that combines off-axis Digital Holography and Dark Field Microscopy to track 100nm gold particles diffusing in water. We show that a single hologram is sufficient to localize several particles in a thick sample with a localization accuracy independent of the particle position. From our measurements we reconstruct the trajectories of the particles and derive their 3D diffusion coefficient. Our results pave the way for quantitative studies of the motion of single nanoparticle in complex media.

This paper describes an imaging microscopic technique based on heterodyne digital holography where subwavelengthsized gold colloids can be imaged in cell environment. Surface cellular receptors of 3T3 mouse fibroblasts are labeled with 40 nm gold nanoparticles, and the biological specimen is imaged in a total internal reflection configuration with holographic microscopy. Due to a higher scattering efficiency of the gold nanoparticles versus that of cellular structures, accurate localization of a gold marker is obtained within a 3D mapping of the entire sample's scattered field, with a lateral precision of 5 nm and 100 nm in the x,y and in the z directions respectively, demonstrating the ability of holographic microscopy to locate nanoparticles in living cells environments.

We obtain quantitative measurements of the oscillation amplitude of vibrating objects by using sideband digital holography. The frequency sidebands on the light scattered by the object, shifted by n times the vibration frequency, are selectively detected by heterodyne holography, and images of the object are calculated for different orders n. Orders up to n = 120 have been observed, allowing the measurement of amplitudes of oscillation that are significantly larger than the optical wavelength. Using the positions of the zeros of intensity for each value of n, we reconstruct the shape of vibration the object.

In the measurement of the amplitude of vibration of objects, holographic imaging techniques usually involve fringe counting; because of the limited resolution of the images, measurements of large amplitudes are not accessible. We demonstrate a technique that suppresses the necessity of fringe counting--frequency sideband imaging--where the order of the sideband is considered a marker of the amplitude. The measurement is completely local: no comparison with another reference point on the object is necessary. It involves a sharp variation of a signal, which makes it robust against perturbations. The method is demonstrated in an experiment made with a vibrating clarinet reed; phase modulations as large as 1000 rad have been measured.