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Description:

Inverted Ratio-fluorescence microscope that is a useful tool to study dynamic events in live cells and cellular structures. Unlike conventional intensity-based fluorescence microscopy, ratio-fluorescence eliminates the effects of photo-bleaching, cell path length variation, non-uniformity of dye loading, the non-uniform field of view, making ratio-fluorescence the most accurate method to determine intracellular ion concentrations for Ca2+, H+ and Na+. The Nikon Eclipse TE2000U microscope is equipped with a xenon arc lamp that provides high intensity, continuous broadband illumination, a monochromatic Hamamatsu camera and high-speed shutters that facilitate the detection of essential dynamic information about ion concentrations and interactions with other molecules, drugs or added reagents. It is ideally suited for measuring changes in intracellular Ca2+ levels, assessed by the ratio of Fura-2 fluorescence at 340 and 380 nm wavelengths. This system also has a 365 nm excitation filter that enables the assessment of Fura-2 intensity level independent of free Ca2+ ion levels, i.e. at the isobestic point of Fura-2.

Illumination:

• Halogen and Xenon Arc Lamp

Objectives:

• 10x/0.25 (PH 1; WD 7.0)

• S FLUOR 40x/1.3 (WD 0.22)

• S FLUOR 100x/0.5-1.3 (WD 0.20)

Cameras:

• Hamamatsu ORCA ER (monochrome)

Filter Cubes: 340 nm, 365 nm, 380 nm and FITC

Software: Simple PCI