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The fluorescence from an open volume of a solution of fluorescent molecules fluctuates as the molecules randomly diffuse into and out of the volume. The distribution of degrees of aggregation or polymerization of the fluorescent molecules can be characterized without perturbing the system by measuring either the moments or the amplitude distribution of these fluctuations. We present an experimental verification of this approach applied to simple model systems consisting of solutions of fluorescent particles of well-defined size. We have also characterized the response of the photon-detection device (typically a photomultiplier), which is essential to the analysis of the fluorescence fluctuations, and have compared two methods for determining shot-noise contributions.

Distribution of molecular aggregation by analysis of fluctuation moments.