Photonics, Vol. 10, Pages 601: Background Rejection in Two-Photon Fluorescence Image Scanning Microscopy

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Photonics, Vol. 10, Pages 601: Background Rejection in Two-Photon Fluorescence Image Scanning Microscopy

Photonics doi: 10.3390/photonics10050601

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

We discuss the properties of signal strength and integrated intensity in two-photon excitation confocal microscopy and image scanning microscopy. The resolution, optical sectioning and background rejection are all improved over nonconfocal two-photon microscopy. Replacing the pinhole of confocal two-photon microscopy with a detector array increases the peak intensity of the point spread function. The outer pixels of a detector array give signals from defocused regions, and thus the processing of these, such as through subtraction, can further improve optical sectioning and background rejection.

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