Special Section on Optical Diagnostic Imaging from Bench to BedsideCombined Imaging

Integrated photothermal flow cytometry in vivo

[+] Author Affiliations
Vladimir P. Zharov

University of Arkansas for Medical Sciences (UAMS), Philips Classic Laser Laboratories, 4301 West Markham St. #543, Little Rock, Arkansas 72205-7199

Ekaterina I. Galanzha

University of Arkansas for Medical Sciences (UAMS), Philips Classic Laser Laboratories, 4301 West Markham St. #543, Little Rock, Arkansas 72205-7199 and Saratov State University, Saratov, Russia 410012

Valery V. Tuchin

Saratov State University, Saratov, Russia 410012

J. Biomed. Opt. 10(5), 051502 (October 31, 2005). doi:10.1117/1.2070167
History: Received December 01, 2004; Revised February 04, 2005; Accepted February 08, 2005; Published October 31, 2005
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The capability of integrated flow cytometry to detect, in real time, moving cells in their natural states in vivo is demonstrated in a study of circulating red and white blood cells in lymph and blood flow of rat mesentery. This system combines dual pump-probe photothermal (PT) techniques, such as PT imaging, the PT thermolens method, and PT velocimetry, with high-resolution (up to 0.3μm), high-speed (up to 1000fps) transmission digital microscopy (TDM) and fluorescence imaging. All PT techniques are based on irradiation of cells in rat mesenteric microvessels with a spectrally tunable laser pulse (420to570nm, 8ns, 0.1to300μJ) and on detection of temperature-dependent variations of the refractive index with a second continuous probe laser beam (633nm, 1.4mW). We focus on intravital monitoring of the integral PT response from single, moving, unlabeled cells (from 100 to 500 cells in one measurement). Potential in vivo applications of this new optical tool, called PT flow cytometry (PTFC), are discussed, including identification of selected cells with differences in natural absorptive properties and sizes, determination of laser-induced cell damage, estimation of flow velocity, and monitoring of circulating cells labeled with PT probes.

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© 2005 Society of Photo-Optical Instrumentation Engineers

Citation

Vladimir P. Zharov ; Ekaterina I. Galanzha and Valery V. Tuchin
"Integrated photothermal flow cytometry in vivo", J. Biomed. Opt. 10(5), 051502 (October 31, 2005). ; http://dx.doi.org/10.1117/1.2070167


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