Fluorescence Ratio Imaging
Fluorescence ratio imaging is the monitoring of live cells in which a fluorescent indicator of intracellular ions is introduced. Indicator dyes have been designed to shift their fluorescence excitation or emission spectrum when binding with specific ions. Images are obtained at two different wavelengths, typically matching the absorption bands at the high and low binding conditions.
By ratioing the intensities in the images, it is possible to construct a map showing the local ion concentrations throughout the field of view. Since the monitoring process is nondestructive, image acquisition can be repeated frequently to trace and monitor the time course of cellular responses.
The MetaFluor® Imaging System is designed for dual-wavelength intracellular ion measurements. The system provides simultaneous display of the raw data, ratio image, graphs of intensities, ratios and ion concentrations, and a non-ratiometric image such as a brightfield or phase-contrast image. Two different ratiometric indicators can be imaged and measured simultaneously.
Toolbars, menus, wizards and dialog boxes help move you through the image processing steps quickly. Features such as multiple image windows, flexible device control, synchronization and timing, and journals allow for automated image acquisition and analysis unlike any other system.
With MetaFluor, you customize the set-up once, then let the experiment run by itself. You are able to collect a large amount of data online and process it with either MetaFluor or an analysis-only copy of the software.