Abstract
In this chapter we will discuss the properties of three-dimensional image formation in an optical fluorescence microscope. An image is blurred and distorted by noise during its formation and acquisition. This distortion hides fine details in the image hampering both the visual and the quantitative analysis of the image. In order to appreciate the fundamental limitations of the resolution and image formation properties of an optical fluorescence microscope, it is necessary to begin by discussing the foundations of diffraction theory. Using this theory, the wave description and the quantum nature of light it is possible to derive a model for the image formation in a general epi-fluorescence microscope and discuss the conditions under which a fluorescence microscope can be modeled as a linear translation invariant system. This means that the optical system is completely described by its Point Spread Function (PSF). The image formations in a Confocal Laser Scanning Microscope (CLSM), in a Wide-Field Microscope and in a Two- Photon Excitation (TPE) Microscope are derived as limited case of the general epi-fluorescence microscope.
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Vicidomini, G. (2005). Image Formation in Fluorescence Microscopy. In: Evangelista, V., Barsanti, L., Passarelli, V., Gualtieri, P. (eds) From Cells to Proteins: Imaging Nature across Dimensions. NATO Security through Science Series. Springer, Dordrecht . https://doi.org/10.1007/1-4020-3616-7_18
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DOI: https://doi.org/10.1007/1-4020-3616-7_18
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