Whole-body imaging with fluorescent proteins

Whole-body imaging with fluorescent proteins
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DOI:
10.1038/nprot.2006.223
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发表时间:
2006-01-01
期刊:
影响因子:
14.8
通讯作者:
Yang, Meng
Yang, Meng
中科院分区:
生物学1区
文献类型:
--
作者:
Hoffman, Robert M.;Yang, Meng

文献摘要

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利用荧光蛋白的固有亮度开发了肿瘤和小鼠内脏器官基因表达的全身成像技术。荧光蛋白基因的稳定转化可以使用含有选择标记如新霉素抗性的逆转录病毒载体来实现。然后可以将稳定表达荧光蛋白的细胞移植到适当的小鼠模型中。对于全身成像,裸鼠非常合适。如果使用野生型小鼠,则必须通过剃毛或脱毛去除毛发。使用的仪器范围从简单的 LED 手电筒和适当的激发和发射滤光片到复杂的设备,例如具有多种放大倍率的 Olympus OV100,可实现宏观成像和微观成像。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基本上所有器官上的荧光细胞的全身成像。这些实验的时间从 2 天到 2 个月不等。
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 retroviral 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. If wild-type mice are used, then hair must be removed by shaving or depilation. The instruments used can range from a simple LED flashlight and appropriate excitation and emission filters to sophisticated equipment such as the Olympus OV100 with a wide range of magnification, enabling both macroimaging and microimaging. 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. The timeline for these experiments varies from 2 days to 2 months.