Background Rejection in Two-Photon Fluorescence Image Scanning Microscopy
Published in Photonics, 2023
Recommended citation: Sheppard, C.J.R.; Castello, M.; Tortarolo, G.; Zunino, A.; Slenders, E.; Bianchini, P.; Vicidomini, G.; Diaspro, A. Background Rejection in Two-Photon Fluorescence Image Scanning Microscopy. Photonics 2023, 10, 601. https://doi.org/10.3390/photonics10050601
Abstract
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.