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基于慢病毒的DsRed-2转染胰腺癌细胞用于转移性疾病的体内深度成像。

Lentivirus-based DsRed-2-transfected pancreatic cancer cells for deep in vivo imaging of metastatic disease.

作者信息

Yu Zeqian, Zhou Jiahua, Hoffman Robert M

机构信息

Department of General Surgery, Zhongda Hospital, Southeast University, Nan Jing City, Jiang Su Province, China.

出版信息

Methods Mol Biol. 2012;872:69-83. doi: 10.1007/978-1-61779-797-2_5.

Abstract

Pancreatic cancer is one of the most aggressive human malignancies. One of the leading causes of pancreatic cancer death is metastasis. The early stages of tumor progression and micrometastasis formation have been difficult to analyze and have been hampered by the inability to identify small numbers of tumor cells against a background of many host cells. The intrinsic brightness of fluorescent proteins has been taken advantage of to develop a technology of whole-body imaging of tumors and gene expression in mouse internal organs. Stable transformation with fluorescent protein genes can be effected using lentiviral vectors containing a selectable marker such as neomycin resistance. The cells that stably express fluorescent proteins can then be transplanted into appropriate mouse models. For whole-body imaging, nude mice are very appropriate, the hair does not need to be removed by shaving or depilation. The instruments used are very simple, they need appropriate excitation and emission filters. It is crucial that proper filters be used such that background autofluorescence is minimal. Fluorescent protein-based imaging technology can be used for whole-body imaging of fluorescent cells on essentially all organs.

摘要

胰腺癌是最具侵袭性的人类恶性肿瘤之一。胰腺癌死亡的主要原因之一是转移。肿瘤进展和微转移形成的早期阶段一直难以分析,并且由于无法在众多宿主细胞的背景中识别少量肿瘤细胞而受到阻碍。荧光蛋白的固有亮度已被用于开发一种对小鼠内脏器官中的肿瘤和基因表达进行全身成像的技术。使用含有如新霉素抗性等选择标记的慢病毒载体可以实现荧光蛋白基因的稳定转化。然后可以将稳定表达荧光蛋白的细胞移植到合适的小鼠模型中。对于全身成像,裸鼠非常合适,不需要通过剃毛或脱毛去除毛发。所使用的仪器非常简单,它们需要合适的激发和发射滤光片。使用适当的滤光片以使背景自发荧光最小化至关重要。基于荧光蛋白的成像技术可用于对基本上所有器官上的荧光细胞进行全身成像。

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