Hodgson Louis, Shen Feimo, Hahn Klaus
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine of Yeshiva University, Gruss-Lipper Biophotonics Center, Bronx, New York, USA.
Curr Protoc Cell Biol. 2010 Mar;Chapter 14:Unit 14.11.1-26. doi: 10.1002/0471143030.cb1411s46.
The biosensors developed in the authors' laboratory have been based on different designs, each imparting specific strengths and weaknesses. Here we describe detailed protocols for the application of three biosensors exemplifying different designs-first, a design in which an environmentally sensitive dye is used to report the activation of endogenous Cdc42, followed by two biosensors based on FRET, one using intramolecular and the other intermolecular FRET. The design differences lead to the need for different approaches in imaging and image analysis.
作者实验室开发的生物传感器基于不同的设计,每种设计都有其特定的优缺点。在此,我们描述了三种体现不同设计的生物传感器应用的详细方案——首先是一种使用环境敏感染料来报告内源性Cdc42激活的设计,接着是两种基于荧光共振能量转移(FRET)的生物传感器,一种使用分子内FRET,另一种使用分子间FRET。设计上的差异导致在成像和图像分析中需要采用不同的方法。