Over the last decade, the light microscope has become increasingly useful as a quantitative tool for studying colloidal systems. The ability to obtain particle coordinates in bulk samples from micrographs is particularly appealing. In this paper we review and extend methods for optimal image formation of colloidal samples, which is vital for particle coordinates of the highest accuracy, and for extracting the most reliable coordinates from these images. We discuss in depth the accuracy of the coordinates, which is sensitive to the details of the colloidal system and the imaging system. Moreover, this accuracy can vary between particles, particularly in dense systems. We introduce a previously unreported error estimate and use it to develop an iterative method for finding particle coordinates. This individual-particle accuracy assessment also allows comparison between particle locations obtained from different experiments. Though aimed primarily at confocal microscopy studies of colloidal systems, the methods outlined here should transfer readily to many other feature extraction problems, especially where features may overlap one another.
Individual colloidal particles have been studied experimentally in a one dimensional random potential with energies that follow a Gaussian distribution. This rough, noise-like potential has been realised using a holographic optical set-up, which allows the width of the distribution to be varied. For different widths, the particle trajectories were followed and the particle dynamics characterised by, for example, the mean square displacement, non-Gaussian parameter, van Hove function, time-dependent diffusion coefficient and residence time distribution. The values obtained for these observables are consistent with the static properties of the system, in particular the barrier height distribution, which was obtained by a detailed characterisation of the tweezer-like set-up. The dynamics display three distinct behaviours: at short times normal diffusion, subsequently an extended regime of localisation within the different minima of the potential and finally a very slow approach towards long-time diffusive behaviour, for which diffusion coefficients consistent with theoretical predictions have been found.
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