Model-based resolution: applying the theory in quantitative microscopy

Model-based image processing techniques have been proposed as a way to increase the resolution of optical microscopes. Here a model based on the microscope's point-spread function is analyzed, and the resolution Limits achieved with a proposed goodness-of-fit criterion are quantified. Several experiments were performed to evaluate the possibilities and limitations of this method: (a) experiments with an ideal (diffraction-limited) microscope, (b) experiments with simulated dots and a real microscope, and (c) experiments with real dots acquired with a real microscope. The results show that a threefold increase over classical resolution (e.g., Rayleigh) is possible. These results can be affected by model misspecifications, whereas model corruption, as seen in the effect of Poisson noise, seems to be unimportant. This research can be considered to he preliminary with the final goal being the accurate measurement of various cytogenetic properties, such as gene distributions, in labeled preparations. (C) 2000 Optical Society of America. OCIS codes: 100.6640. 170.2520, 110.4280, 110.0180, 120.4640, 100.2960.