Sankaran Jagadish, Shi Xianke, Ho Liang Yoong, Stelzer Ernst H K, Wohland Thorsten
Singapore-MIT Alliance, National University of Singapore, E4-04-10, 4 Engineering Drive 3, 117576 Singapore.
Opt Express. 2010 Dec 6;18(25):25468-81. doi: 10.1364/OE.18.025468.
The multiplexing of fluorescence correlation spectroscopy (FCS), especially in imaging FCS using fast, sensitive array detectors, requires the handling of large amounts of data. One can easily collect in excess of 100,000 FCS curves a day, too many to be treated manually. Therefore, ImFCS, an open-source software which relies on standard image files was developed and provides a wide range of options for the calculation of spatial and temporal auto- and cross-correlations, as well as differences in Cross-Correlation Functions (ΔCCF). ImFCS permits fitting of standard models to correlation functions and provides optimized histograms of fitted parameters. Applications include the measurement of diffusion and flow with Imaging Total Internal Reflection FCS (ITIR-FCS) and Single Plane Illumination Microscopy FCS (SPIM-FCS) in biologically relevant samples. As a compromise between ITIR-FCS and SPIM-FCS, we extend the applications to Imaging Variable Angle-FCS (IVA-FCS) where sub-critical oblique illumination provides sample sectioning close to the cover slide.
荧光相关光谱（FCS）的多路复用，特别是在使用快速、灵敏的阵列探测器进行成像FCS时，需要处理大量数据。一天内轻松就能收集超过10万条FCS曲线，数量太多无法手动处理。因此，开发了ImFCS，这是一款依赖标准图像文件的开源软件，它为空间和时间自相关及互相关的计算，以及互相关函数差异（ΔCCF）提供了广泛的选项。ImFCS允许将标准模型拟合到相关函数，并提供拟合参数的优化直方图。应用包括在生物相关样本中使用成像全内反射FCS（ITIR-FCS）和单平面照明显微镜FCS（SPIM-FCS）测量扩散和流动。作为ITIR-FCS和SPIM-FCS之间的一种折中，我们将应用扩展到成像可变角度FCS（IVA-FCS），其中亚临界斜照明可在盖玻片附近提供样品切片。
