Photoacoustic probing of fluorophore excited state lifetime with application to oxygen sensing

• Published in: Journal of biomedical optics Vol. 13; no. 3; p. 034023
• Main Authors: Ashkenazi, Shai, Huang, Shen-Wen, Horvath, Thomas, Koo, Yong-Eun L, Kopelman, Raoul, Kopelman, Raowl
• Format: Journal Article
• Language: English
• Published: United States 01.05.2008
• Subjects: Acoustics; Computer Simulation; Fluorescence Resonance Energy Transfer - methods; Fluorescent Dyes - analysis; Fluorescent Dyes - chemistry; Lasers; Models, Chemical; Molecular Probe Techniques; Nephelometry and Turbidimetry - methods; Oximetry - methods; Photochemistry - methods
• ISSN: 1083-3668
• DOI: 10.1117/1.2927466

A new method is developed to perform local measurements of fluorophore excited state lifetimes in turbid media without collecting the fluorescence emission. The method is based on a pump-probe approach where a first laser pulse excites the dye and then a second laser pulse is used for photoacoustic probing of the transient absorption. The photoacoustic response generated by the probe pulse is recorded by an ultrasound receiver. Repeating the measurement for increasing pump-probe time delays yields a series of photoacoustic signals that are used to extract the lifetime of the excited state. The method is validated by measuring the lifetime of an oxygen sensitive dye solution at different concentrations of dissolved oxygen. The dye is pumped with a 532-nm pulsed laser and the transient absorption at 740 nm is probed using a second pulsed laser system. The photoacoustic-based results are in close agreement with those obtained from time-dependent fluorescent measurements. The method can be extended to photoacoustic lifetime imaging by using a receiver array instead of a single receiver. Potential applications of this method include tissue oxygen imaging for cancer diagnostics and mapping molecular events such as resonant energy transfer and ion collisions in a biological environment.