Weisshart Klaus, Jüngel Volker, Briddon Stephen J
Carl Zeiss Jena GmbH, Carl-Zeiss-Promenade 10, 07745 Jena, Germany.
Curr Pharm Biotechnol. 2004 Apr;5(2):135-54. doi: 10.2174/1389201043376913.
Fluorescence Correlation Spectroscopy (FCS) has developed into a routine method to quantitatively study diffusion of molecules and kinetic processes. FCS has recently become popular to complement live cell imaging with biophysical information. The enabling technology has been commercially realised by combining laser scanning microscopes and fluorescence correlation spectrometers in one integrated platform. This article provides an overview of the Zeiss solution of such a combined instrument, the LSM 510 META / ConfoCor 2 system. We focus on the instrumental set up as well as technical advances and improvements in the software that controls FCS data acquisition and evaluation. In addition, we outline the calibration of the instrument and the work flow for data analysis with emphasis on in vivo pharmacological applications.
荧光相关光谱法(FCS)已发展成为一种用于定量研究分子扩散和动力学过程的常规方法。最近,FCS因能为活细胞成像补充生物物理信息而受到欢迎。通过将激光扫描显微镜和荧光相关光谱仪整合在一个平台上,这项技术已实现商业化。本文概述了蔡司公司的此类组合仪器解决方案,即LSM 510 META / ConfoCor 2系统。我们重点关注仪器设置以及控制FCS数据采集和评估的软件方面的技术进步和改进。此外,我们还概述了仪器校准和数据分析工作流程,重点是体内药理学应用。