title: Fiber-optic multiphoton flow cytometry in whole blood and in vivo.
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.
authors.affiliation: National Changhua University of Education, Department of Electrical Engineering, Changhua, Taiwan. ycchang@cc.ncue.edu.tw
authors.forename: Yu-Chung
authors.initials: YC
authors.lastname: Chang
authors.forename: Jing Yong
authors.initials: JY
authors.lastname: Ye
authors.forename: Thommey P
authors.initials: TP
authors.lastname: Thomas
authors.forename: Zhengyi
authors.initials: Z
authors.lastname: Cao
authors.forename: Alina
authors.initials: A
authors.lastname: Kotlyar
authors.forename: Eric R
authors.initials: ER
authors.lastname: Tkaczyk
authors.forename: James R
authors.initials: JR
authors.lastname: Baker
authors.forename: Theodore B
authors.initials: TB
authors.lastname: Norris
mesh_terms: D000818:Animals; D058948:Cell Tracking / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005336:Fiber Optic Technology / Q000295:instrumentation*; D005434:Flow Cytometry / Q000295:instrumentation*; D006801:Humans; D051379:Mice; D036641:Microscopy, Fluorescence, Multiphoton / Q000295:instrumentation*; D009360:Neoplastic Cells, Circulating / Q000473:pathology*
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