TE200 ratiometric imaging (Intracellular Calcium and pH imaging):
This microscope is heavily used for intracellular calcium measurement experiments with Fura2, however it can be used for other intracellular measurements such as pH, magnesium, Zinc, membrane potential, reactive oxygen species, etc using ratiometric and non ratiometric fluorescent sensor dyes. This system is intended for short term (less than 2hrs) live cell experiments involving fast image capture with sub-second to a few second intervals with easy access to the sample for experimental manipulation such us pharmacological stimulation series or solution composition changes. This microscope uses open-style stage-top incubators with optional multi-channel perfusion. The system is configured with a Nikon TE200 inverted epifluorescence microscope with phase contrast and DIC capabilities, excitation is provided by a Sutter Instruments 150 watt Xenon DG4 illuminator capable of 2 ms changes in up to 4 excitation wavelengths. Image capture is performed by a Roper Scientific CoolSnap HQ 12 bit cooled monochrome CCD camera and instrumentation control are provided by MetaFluor and Metamoprh software (Molecular Devices).