1 July 2010 Fiber-optic multiphoton flow cytometry in whole blood and in vivo
Author Affiliations +
J. of Biomedical Optics, 15(4), 047004 (2010). doi:10.1117/1.3463481
Abstract
Circulating tumor cells in the bloodstream are sensitive indicators for metastasis and disease prognosis. Circulating cells have usually been monitored via extraction from blood, and more recently in vivo using free-space optics; however, long-term intravital monitoring of rare circulating cells remains a major challenge. We demonstrate the application of a two-photon-fluorescence optical fiber probe for the detection of cells in whole blood and in vivo. A double-clad fiber was used to enhance the detection sensitivity. Two-channel detection was employed to enable simultaneous measurement of multiple fluorescent markers. Because the fiber probe circumvents scattering and absorption from whole blood, the detected signal strength from fluorescent cells was found to be similar in phosphate-buffered saline (PBS) and in whole blood. The detection efficiency of cells labeled with the membrane-binding dye 1,1-dioctadecyl-3,3,3,3-tetramethylindoldicarbocyanine, 4-chlorobenzenesulfonate (DiD) was demonstrated to be the same in PBS and in whole blood. A high detection efficiency of green fluorescent protein (GFP)-expressing cells in whole blood was also demonstrated. To characterize in vivo detection, DiD-labeled untransfected and GFP-transfected cells were injected into live mice, and the cell circulation dynamics was monitored in real time. The detection efficiency of GFP-expressing cells in vivo was consistent with that observed ex vivo in whole blood.
Yu-Chung Chang, Jing Yong Ye, Thommey P. Thomas, Zhengyi Cao, Alina Kotlyar, Eric R. Tkaczyk, James R. Baker, Theodore B. Norris, "Fiber-optic multiphoton flow cytometry in whole blood and in vivo," Journal of Biomedical Optics 15(4), 047004 (1 July 2010). http://dx.doi.org/10.1117/1.3463481
Submission: Received ; Accepted
JOURNAL ARTICLE
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KEYWORDS
Blood

In vivo imaging

Green fluorescent protein

Luminescence

Flow cytometry

Photon counting

Fiber optics

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