Multimodal wide-field two-photon excitation imaging: characterization of the technique for in vivo applications
Abstract
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.
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.Attached Files
Published - Hwang2011p17346Biomed_Opt_Express.pdf
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Additional details
- PMCID
- PMC3038450
- Eprint ID
- 29586
- Resolver ID
- CaltechAUTHORS:20120306-091219207
- Navy Bureau of Medicine and Surgery
- Caltech-City of Hope Biomedical Initiative
- Created
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2012-03-13Created from EPrint's datestamp field
- Updated
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2021-11-09Created from EPrint's last_modified field