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Multimodal wide-field two-photon excitation imaging: characterization of the technique for in vivo applications

Hwang, Jae Youn and Wachsmann-Hogiu, Sebastian and Ramanujan, V. Krishnan and Nowatzyk, Andreas G. and Koronyo, Yosef and Medina-Kauwe, Lali K. and Gross, Zeev and Gray, Harry B. and Farkas, Daniel L. (2011) Multimodal wide-field two-photon excitation imaging: characterization of the technique for in vivo applications. Biomedical Optics Express, 2 (2). pp. 356-364. ISSN 2156-7085. PMCID PMC3038450.

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We report fast, non-scanning, wide-field two-photon fluorescence excitation with spectral and lifetime detection for in vivo biomedical applications. We determined the optical characteristics of the technique, developed a Gaussian flat-field correction method to reduce artifacts resulting from non-uniform excitation such that contrast is enhanced, and showed that it can be used for ex vivo and in vivo cellular-level imaging. Two applications were demonstrated: (i) ex vivo measurements of beta-amyloid plaques in retinas of transgenic mice, and (ii) in vivo imaging of sulfonated gallium(III) corroles injected into tumors. We demonstrate that wide-field two photon fluorescence excitation with flat-field correction provides more penetration depth as well as better contrast and axial resolution than the corresponding one-photon wide field excitation for the same dye. Importantly, when this technique is used together with spectral and fluorescence lifetime detection modules, it offers improved discrimination between fluorescence from molecules of interest and autofluorescence, with higher sensitivity and specificity for in vivo applications.

Item Type:Article
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URLURL TypeDescription CentralUNSPECIFIED
Gray, Harry B.0000-0002-7937-7876
Additional Information:© 2011 Optical Society of America. Received 9 Dec 2010; revised 9 Jan 2011; accepted 10 Jan 2011; published 13 Jan 2011. Partial support from the U.S. Navy Bureau of Medicine and Surgery is gratefully acknowledged. Work at Caltech was supported by a CIT-COH initiative grant.
Funding AgencyGrant Number
Navy Bureau of Medicine and SurgeryUNSPECIFIED
Caltech-City of Hope Biomedical InitiativeUNSPECIFIED
Issue or Number:2
Classification Code:OCIS codes: 180.4315, 110.4234
PubMed Central ID:PMC3038450
Record Number:CaltechAUTHORS:20120306-091219207
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Official Citation:Jae Youn Hwang, Sebastian Wachsmann-Hogiu, V Krishnan Ramanujan, Andreas G. Nowatzyk, Yosef Koronyo, Lali K. Medina-Kauwe, Zeev Gross, Harry B. Gray, and Daniel L. Farkas, "Multimodal wide-field two-photon excitation imaging: characterization of the technique for in vivo applications," Biomed. Opt. Express 2, 356-364 (2011)
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:29586
Deposited By: Ruth Sustaita
Deposited On:13 Mar 2012 18:16
Last Modified:22 Nov 2019 09:58

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