Fiber-optic multiphoton flow cytometry in whole blood and in vivo
Fiber-optic multiphoton flow cytometry in whole blood and in vivo
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DOI：
10.1117/1.3463481
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发表时间：
2010-07-01
影响因子：
3.5
通讯作者：
Norris, Theodore B.
中科院分区：
文献类型：
--
作者：
Chang, Yu-Chung;Ye, Jing Yong;Norris, Theodore B.
血液循环中的肿瘤细胞是肿瘤转移和疾病预后的敏感指标。循环细胞通常通过从血液中提取来监测，最近在体内使用自由空间光学；然而，罕见循环细胞的长期生命监测仍然是一个重大挑战。我们展示了一种双光子荧光光纤探针在全血和体内细胞检测中的应用。采用双包层光纤提高了探测灵敏度。采用双通道检测，可同时检测多种荧光标记物。由于纤维探针避开了全血的散射和吸收，从荧光细胞中检测到的信号强度在磷酸盐缓冲盐水（PBS）和全血中相似。用膜结合染料1,1'-二十八烷基-3,3,3',3'-四甲基吲哚二碳菁，4-氯苯磺酸（DiD）标记的细胞在PBS和全血中的检测效率相同。全血中表达绿色荧光蛋白（GFP）的细胞具有较高的检测效率。为了表征体内检测，将未转染的did标记细胞和转染gfp的细胞注射到活小鼠体内，并实时监测细胞循环动力学。体内表达gfp的细胞检测效率与体外全血检测效率一致。(C) 2010光学仪器工程师学会。(DOI: 10.1117/1.3463481)
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. (C) 2010 Society of Photo-Optical Instrumentation Engineers. [DOI: 10.1117/1.3463481]