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Signal strength and integrated intensity in confocal and image scanning microscopy

Colin J. R. Sheppard, Marco Castello, Giorgio Tortarolo, Alessandro Zunino, Eli Slenders, Paolo Bianchini, Giuseppe Vicidomini, and Alberto Diaspro

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Abstract

The properties of signal strength and integrated intensity in a scanned imaging system are reviewed. These properties are especially applied to confocal imaging systems, including image scanning microscopy. The integrated intensity, equal to the image of a uniform planar (sheet) object, rather than the peak of the point spread function, is a measure of the flux in an image. Analytic expressions are presented for the intensity in the detector plane for a uniform volume object, and for the resulting background. The variation in the integrated intensity with defocus for an offset point detector is presented. This axial fingerprint is independent of any pixel reassignment. The intensity in the detector plane is shown to contain the defocus information, and simple processing of the recorded data can improve optical sectioning and background rejection.

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Data underlying the results presented in this paper are not publicly available at this time but may be obtained from the authors upon reasonable request.

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