2 resultados para light microscopy

em DI-fusion - The institutional repository of Université Libre de Bruxelles


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Whereas the resolving power of an ordinary optical microscope is determined by the classical Rayleigh distance, significant super-resolution, i.e. resolution improvement beyond that Rayleigh limit, has been achieved by confocal scanning light microscopy. Furthermore is has been shown that the resolution of a confocal scanning microscope can still be significantly enhanced by measuring, for each scanning position, the full diffraction image by means of an array of detectors and by inverting these data to recover the value of the object at the focus. We discuss the associated inverse problem and show how to generalize the data inversion procedure by allowing, for reconstructing the object at a given point, to make use also of the diffraction images recorded at other scanning positions. This leads us to a whole family of generalized inversion formulae, which contains as special cases some previously known formulae. We also show how these exact inversion formulae can be implemented in practice.

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For pt.I see ibid. vol.3, p.195 (1987). The authors have shown that the resolution of a confocal scanning microscope can be improved by recording the full image at each scanning point and then inverting the data. These analyses were restricted to the case of coherent illumination. They investigate, along similar lines, the incoherent case, which applies to fluorescence microscopy. They investigate the one-dimensional and two-dimensional square-pupil problems and they prove, by means of numerical computations of the singular value spectrum and of the impulse response function, that for a signal-to-noise ratio of, say 10%, it is possible to obtain an improvement of approximately 60% in resolution with respect to the conventional incoherent light confocal microscope. This represents a working bandwidth of 3.5 times the Rayleigh limit.