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In this paper, we present a maximum likelihood (ML) approach to high-resolution estimation of the shifts of a spectral signal. This spectral signal arises in application of optically based resonant biosensors, where high resolution in the estimation of signal shift is synonymous with high sensitivity to biological interactions. For the particular sensor of interest, the underlying signal is nonuniformly sampled and exhibits Poisson amplitude statistics. Shift estimation accuracies orders of magnitude finer than the sample spacing are sought. The new ML-based formulation leads to a solution approach different from typical resonance shift estimation methods based on polynomial fitting and peak (or ) estimation and tracking.