Hoffman Robert M, Yang Meng
AntiCancer, Inc., 7917 Ostrow Street, San Diego, California 92111, USA.
Nat Protoc. 2006;1(3):1429-38. doi: 10.1038/nprot.2006.223.
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.
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逆转录病毒载体,可以实现用荧光蛋白基因进行稳定转化。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。如果使用野生型小鼠,则必须通过剃毛或脱毛去除毛发。所使用的仪器范围很广,从简单的LED手电筒和合适的激发和发射滤光片到复杂的设备,如具有广泛放大倍数的奥林巴斯OV100,既可以进行宏观成像也可以进行微观成像。使用合适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。这些实验的时间跨度从2天到2个月不等。