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小鼠肿瘤模型的定量生物发光成像

Quantitative bioluminescence imaging of mouse tumor models.

作者信息

Tseng Jen-Chieh, Kung Andrew L

机构信息

Lurie Family Imaging Center, Dana-Farber Cancer Institute, Boston, Massachusetts 02215.

Department of Pediatrics, Columbia University Medical Center, New York, New York 10032.

出版信息

Cold Spring Harb Protoc. 2015 Jan 5;2015(1):pdb.prot078261. doi: 10.1101/pdb.prot078261.

Abstract

Bioluminescence imaging (BLI) has become an essential technique for preclinical evaluation of anticancer therapeutics and provides sensitive and quantitative measurements of tumor burden in experimental cancer models. For light generation, a vector encoding firefly luciferase is introduced into human cancer cells that are grown as tumor xenografts in immunocompromised hosts, and the enzyme substrate luciferin is injected into the host. Alternatively, the reporter gene can be expressed in genetically engineered mouse models to determine the onset and progression of disease. In addition to expression of an ectopic luciferase enzyme, bioluminescence requires oxygen and ATP, thus only viable luciferase-expressing cells or tissues are capable of producing bioluminescence signals. Here, we summarize a BLI protocol that takes advantage of advances in hardware, especially the cooled charge-coupled device camera, to enable detection of bioluminescence in living animals with high sensitivity and a large dynamic range.

摘要

生物发光成像(BLI)已成为抗癌治疗临床前评估的一项重要技术,并能对实验性癌症模型中的肿瘤负荷进行灵敏且定量的测量。为了产生光,将编码萤火虫荧光素酶的载体导入人类癌细胞中,这些细胞在免疫缺陷宿主中作为肿瘤异种移植物生长,然后将酶底物荧光素注射到宿主体内。或者,报告基因可以在基因工程小鼠模型中表达,以确定疾病的发生和进展。除了异位荧光素酶的表达外,生物发光还需要氧气和ATP,因此只有存活的表达荧光素酶的细胞或组织才能产生生物发光信号。在此,我们总结了一种BLI方案,该方案利用了硬件方面的进展,特别是冷却电荷耦合器件相机,能够在活体动物中以高灵敏度和大动态范围检测生物发光。

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