Background Rejection in Two-Photon Fluorescence Image Scanning Microscopy-Reference-Cited by-同舟云学术

Background Rejection in Two-Photon Fluorescence Image Scanning Microscopy

Published:2023-05-22 Issue:5 Volume:10 Page:601
ISSN:2304-6732
Container-title:Photonics
language:en
Short-container-title:Photonics

Author:

Sheppard Colin J. R.¹²^ORCID,Castello Marco³^ORCID,Tortarolo Giorgio³⁴,Zunino Alessandro³^ORCID,Slenders Eli³,Bianchini Paolo¹^ORCID,Vicidomini Giuseppe³,Diaspro Alberto¹⁵^ORCID

Affiliation:

1. Nanoscopy and NIC@IIT, Istituto Italiano di Tecnologia, Via Enrico Melen, 83 Edificio B, 16152 Genova, Italy

2. Molecular Horizons, School of Chemistry and Molecular Biosciences, University of Wollongong, Wollongong, NSW 2522, Australia

3. Molecular Microscopy and Spectroscopy, Istituto Italiano di Tecnologia, Via Enrico Melen, 83 Edificio B, 16152 Genova, Italy

4. Laboratory of Experimental Biophysics, EPFL, CH-1015 Lausanne, Switzerland

5. Dipartimento di Fisica, University of Genoa, 16145 Genova, Italy

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.

Publisher

MDPI AG

Subject

Radiology, Nuclear Medicine and imaging,Instrumentation,Atomic and Molecular Physics, and Optics

Link

https://www.mdpi.com/2304-6732/10/5/601/pdf

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