Full systems considerations for multiphoton microscopy

David L. Wokosin; John M. Girkin

doi:10.1117/12.424558

24 April 2001 Full systems considerations for multiphoton microscopy

David L. Wokosin, John M. Girkin

Proceedings Volume 4262, Multiphoton Microscopy in the Biomedical Sciences; (2001) https://doi.org/10.1117/12.424558
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

Abstract

A most-important-variables analysis of practical, successful multiphoton excitation fluorescence microscopy is presented. The key strength of multiphoton imaging -- localization of the excitation volume -- helps to decouple the excitation from the emission; the emission no longer needs to be imaged. The presentation starts with maximizing the detected signal and proceeds to the laser source considerations. The main goal is to match the instrumentation to the biological problem. The main aspects covered are... Emission collection: potential signal; Focal plane mismatch: chromatic aberration; Light detected: actual signal; Live cell imaging: preservation of biological function; Deep imaging: preservation of image contrast; Signal production: potential limitations.

Citation Download Citation

David L. Wokosin and John M. Girkin "Full systems considerations for multiphoton microscopy", Proc. SPIE 4262, Multiphoton Microscopy in the Biomedical Sciences, (24 April 2001); https://doi.org/10.1117/12.424558

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KEYWORDS

Signal detection

Lenses

Imaging systems

Objectives

Confocal microscopy

Multiphoton microscopy

Luminescence

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