A comparison of the emission efficiency of four common green fluorescence dyes after internalization into cancer cells
A comparison of the emission efficiency of four common green fluorescence dyes after internalization into cancer cells
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DOI：
10.1021/bc0601626
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发表时间：
2006-11-15
影响因子：
4.7
通讯作者：
Kobayashi, Hisataka
中科院分区：
文献类型：
--
作者：
Hama, Yukihiro;Urano, Yasuteru;Kobayashi, Hisataka
在内窥镜检查或手术期间增强癌症检测的体内光学成像需要具有高发射效率和高信号背景比的靶向荧光探针。准确检测癌症的一种策略是让荧光团内化到癌细胞内，从而允许未结合的荧光团被洗掉或吸收。必须仔细考虑用于此任务的荧光团的选择。对于穿透深度，近红外探针通常是优选的，但其量子效率相对较低。虽然绿色荧光蛋白已广泛用于对小鼠内脏器官的肿瘤进行成像，但由于绿光在组织中的穿透距离短且信号产生效率高，绿色荧光探针更适合对浅表组织进行成像。虽然绿色荧光团的荧光特性在体外是众所周知的，但一旦它们被内化到细胞内，人们对其荧光的关注却很少。在本研究中，比较了与亲和素缀合的四种常见绿色荧光团（Av-荧光素、Av-俄勒冈绿、Av-BODIPY-FL和Av-罗丹明绿）与SHIN3卵巢癌细胞一起孵育后的细胞内化后的发射效率。使用凝集素结合受体系统，亲和素-荧光团缀合物被内吞，并用荧光显微镜和流式细胞术评估它们的荧光。虽然荧光素在细胞外表现出最高的信号，但在四种荧光团中，内在的 Av-罗丹明绿在体外和体内从 SHIN3 卵巢癌细胞中发出最多的光。内化的AV-罗丹明绿复合物似乎定位于内质囊泡。因此，在四种常见的绿色荧光染料中，罗丹明绿是细胞内化后最亮的绿色荧光探针。该信息可能对依赖细胞内化进行癌症检测的肿瘤靶向荧光探针的设计产生影响。
In vivo optical imaging to enhance the detection of cancer during endoscopy or surgery requires a targeted fluorescent probe with high emission efficiency and high signal-to-background ratio. One strategy to accurately detect cancers is to have the fluorophore internalize within the cancer cells permitting nonbound fluorophores to be washed away or absorbed. The choice of fluorophores for this task must be carefully considered. For depth of penetration, near-infrared probes are ordinarily preferred but suffer from relatively low quantum efficiency. Although green fluorescent protein has been widely used to image tumors on internal organs in mice, green fluorescent probes are better suited for imaging the superficial tissues because of the short penetration distance of green light in tissue and the highly efficient production of signal. While the fluorescence properties of green fluorophores are well-known in vitro, less attention has been paid to their fluorescence once they are internalized within cells. In this study, the emission efficiency after cellular internalization of four common green fluorophores conjugated to avidin ( Av-fluorescein, Av-Oregon green, Av-BODIPY-FL, and Av-rhodamine green) were compared after each conjugate was incubated with SHIN3 ovarian cancer cells. Using the lectin binding receptor system, the avidin-fluorophore conjugates were endocytosed, and their fluorescence was evaluated with fluorescence microscopy and flow cytometry. While fluorescein demonstrated the highest signal outside the cell, among the four fluorophores, internalized Av-rhodamine green emitted the most light from SHIN3 ovarian cancer cells both in vitro and in vivo. The internalized Av-rhodamine green complex appeared to localize to the endoplasmic vesicles. Thus, among the four common green fluorescent dyes, rhodamine green is the brightest green fluorescence probe after cellular internalization. This information could have implications for the design of tumor-targeted fluorescent probes that rely on cellular internalization for cancer detection.