Heuvelman Gerrit, Erdel Fabian, Wachsmuth Malte, Rippe Karsten
Research Group Genome Organization and Function, Deutsches Krebsforschungszentrum and BioQuant, Heidelberg, Germany.
Eur Biophys J. 2009 Jul;38(6):813-28. doi: 10.1007/s00249-009-0499-9. Epub 2009 Jun 19.
The spatial and temporal fluctuation microscope (STFM) presented here extends the concept of a fluorescence confocal laser scanning microscope to illumination and detection along a line. The parallel multichannel acquisition of the fluorescence signal was accomplished by using a single line of an electron-multiplying charge-coupled device camera at 14 mus time resolution for detection of the fluorescence signal. The STFM system provided fast confocal imaging (30 images per second) and allowed for the spatially resolved detection of particle concentration fluctuations in fluorescence correlation spectroscopy experiments. For the application of the STFM, an approximated theoretical description of the beam geometry, the point-spread function, and the fluorescence auto- and cross-correlation functions were derived. The STFM was applied to studies of the dynamics of promyelocytic leukemia nuclear bodies, green fluorescent protein, and chromatin-remodeling complexes in living cells. The results demonstrate the unique capabilities of the STFM for characterizing the position-dependent translocations and interactions of proteins in the cell.
本文介绍的空间和时间波动显微镜（STFM）将荧光共聚焦激光扫描显微镜的概念扩展到沿一条线的照明和检测。通过使用电子倍增电荷耦合器件相机的单个线以14微秒的时间分辨率并行多通道采集荧光信号，用于检测荧光信号。STFM系统提供了快速共聚焦成像（每秒30幅图像），并允许在荧光相关光谱实验中对颗粒浓度波动进行空间分辨检测。为了应用STFM，推导了光束几何形状、点扩散函数以及荧光自相关和互相关函数的近似理论描述。STFM被应用于研究活细胞中早幼粒细胞白血病核体、绿色荧光蛋白和染色质重塑复合物的动力学。结果证明了STFM在表征细胞中蛋白质的位置依赖性易位和相互作用方面的独特能力。
