iRFP is a sensitive marker for cell number and tumor growth in high-throughput systems
iRFP is a sensitive marker for cell number and tumor growth in high-throughput systems
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DOI：
10.4161/cc.26985
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发表时间：
2014-01-15
期刊：
影响因子：
4.3
通讯作者：
Vousden, Karen H.
中科院分区：
文献类型：
--
作者：
Hock, Andreas K.;Lee, Pearl;Vousden, Karen H.
GFP和荧光素酶被广泛用作体外和体内的标记物，尽管两者都有局限性。GFP荧光的实用性受到高背景信号和差组织透射率的限制。荧光素酶的通量在体外受到细胞裂解要求的限制，而在体内，荧光素酶读出由于需要底物注射和对内源性ATP的依赖而变得复杂。在这里，我们表明，近红外荧光蛋白与广泛使用的近红外扫描仪相结合，克服了这些障碍，并允许准确测定细胞数量在体外和肿瘤生长在体内以高通量的方式，在可以忽略不计的每孔成本。该系统代表了在跟踪组织培养以及动物中的细胞增殖方面的重大进展，在细胞生物学中具有广泛的应用。
GFP and luciferase are used extensively as markers both in vitro and in vivo although both have limitations. The utility of GFP fluorescence is restricted by high background signal and poor tissue penetrance. Luciferase throughput is limited in vitro by the requirement for cell lysis, while in vivo, luciferase readout is complicated by the need for substrate injection and the dependence on endogenous ATP. Here we show that near-infrared fluorescent protein in combination with widely available near-infrared scanners overcomes these obstacles and allows for the accurate determination of cell number in vitro and tumor growth in vivo in a high-throughput manner and at negligible per-well costs. This system represents a significant advance in tracking cell proliferation in tissue culture as well as in animals, with widespread applications in cell biology.