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The optional diffusion measurement package for the FLUOVIEW FV1200 confocal laser scanning microscope enables data acquisition and the analysis of molecular interactions and concentrations by calculating the diffusion coefficients of molecules within a cell. The range of analytical tools presented by the software includes raster imaging correlation spectroscopy, point fluorescence correlation spectroscopy, cross-correlation spectroscopy and FRAP analysis. This versatility covers a wide range of molecular sizes and speeds for diverse research needs.
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Raster image correlation spectroscopy (RICS) is a method of analyzing the diffusion and binding dynamics of molecules in an entire, single image. RICS uses a spatial correlation algorithm to calculate diffusion coefficients and the number of molecules in specified regions. Cross-correlation RICS (ccRICS) characterizes molecular interactions using fluorescent labelled molecules in two Colors.
Point FCS and point FCCS (point scan fluorescence cross-correlation spectroscopy) can now be performed with the built-in PMTs of the FLUOVIEW FV1200. Point FCS analyses intensity fluctuations caused by diffusion or the binding/unbinding interactions of a protein complex, while point FCCS analyses the fluctuation of fluorescent labelled molecules in two Colors to show when proteins are part of the same complex.
The Axelrod analytical algorithm is installed as a FRAP (fluorescence recovery after photobleaching) analysis method. The algorithm is used to calculate diffusion coefficients and the proportions of diffusing molecules; it thus complements the unique Tornado scan, which photobleaches a circular area.
This unit is fully compatible with the FLUOVIEW FV1000 and FV1000MPE systems.
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