A dynamic view of cellular processes by in vivo fluorescence auto- and cross-correlation spectroscopy
A dynamic view of cellular processes by in vivo fluorescence auto- and cross-correlation spectroscopy
复制标题
DOI：
10.1016/s1046-2023(02)00291-8
复制
发表时间：
2003-01-01
期刊：
影响因子：
4.8
通讯作者：
Schwille, P
中科院分区：
文献类型：
--
作者：
Bacia, K;Schwille, P
荧光相关光谱（FCS）作为一种旨在用生物物理信息补充活细胞图像的技术越来越受欢迎。本文提供了一个简短的概述最近的细胞内FCS的应用和研究人员，谁正在寻求整合FCS到活细胞成像，以获得有关粒子的流动性，局部浓度和分子相互作用的信息的实用指南。简要介绍了FCS的原理，特别强调了实际方面，如选择适当的染料和定位的测量体积在样品中。从自相关曲线中提取参数的可能性和局限性进行了讨论，并提请注意潜在的文物，如光漂白和探针聚集。双色互相关的原理是审查沿着考虑适当的设置和调整。被认为是非理想条件下，包括不完全的焦点重叠和光谱串扰的实际影响。最近的例子都自动和互相关的应用程序证明了FCS的细胞生物学的潜力。(C)2002 Elsevier Science（美国）。All rights reserved.
Fluorescence correlation spectroscopy (FCS) is becoming increasingly popular as a technique that aims at complementing live cell images with biophysical information. This article provides both a short overview over recent intracellular FCS applications and a practical guide for investigators, who are seeking to integrate FCS into live cell imaging to obtain information on particle mobility, local concentrations, and molecular interactions. A brief introduction to the principles of FCS is provided, particularly emphasizing practical aspects such as the choice of appropriate dyes and positioning of the measurement volume in the sample. Possibilities and limitations in extracting parameters from autocorrelation curves are discussed, and attention is drawn to potential artifacts, such as photobleaching and probe aggregation. The principle of dual-color cross-correlation is reviewed along with considerations for proper setup and adjustment. Practical implications of nonideal conditions including incomplete focus overlap and spectral crosstalk are considered. Recent examples of both auto- and cross-correlation applications demonstrate the potential of FCS for cell biology. (C) 2002 Elsevier Science (USA). All rights reserved.