Interactive Tutorials - Spinning Disk Fundamentals

Interactive Tutorials

Wavelength Selection

Matching Filter Sets with Microscope Light Sources

In fluorescence microscopy, the excitation light is generally passed through a bandpass interference filter to select a specific band of wavelengths that are used to illuminate the fluorophore. Depending upon the filter characteristics and the light source, the amount of light available for excitation can vary by a wide margin. This interactive tutorial is designed to enable the visitor to choose between various ZEISS filter sets and common microscope illumination sources to determine the optimum combination for a specific application.

The emission profiles of mercury and metal halide arc discharge lamps are distinct from those of the xenon and LED sources in that several prominent emission lines are present in the ultraviolet, blue, green and yellow spectral regions. These spectral lines are significantly brighter (up to 100 times) than the continuous background. When selecting filter sets, those excitation filters that overlap a spectral line will produce a greater level of excitation. In selecting filters to match LED spectral profiles, those diodes that best overlap the filter bandpass are highlighted on the left-hand side of the tutorial window.

Contributing Authors

Michael W. Davidson - National High Magnetic Field Laboratory, 1800 East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310.