Practical Aspects of Fluorescence Filter Combinations

Section Overview:

The terminology applied to fluorescence filters has become a jumble as a result of various initials utilized by different manufacturers to identify their filters. In this section, we attempt to make some order of this confusing terminology and specifically address how the various filter combinations can be utilized with specific fluorophores. Basically there are three categories of filters to be sorted out: exciter filters, barrier filters and dichromatic beamsplitters (dichroic mirrors), which are usually combined to produce a filter cube or block similar to the one illustrated in Figure 1. Proper selection of filters is the key to successful fluorescence microscopy.

Interference Filters

The following sections contain links to review articles and interactive Java tutorials describing a variety of fluorescence filter combinations from the major manufacturers. Additional information is provided with regards to illumination excitation balancers, selection of fluorophores, and matching specific probes with filter combinations.

Review Articles

Interactive Java Tutorials

Fluorochrome and Fluorescence Filter Combination Data Tables

In this section, we have compiled a listing of data on the most popular fluorochromes including excitation and emission characteristics, suggested cube utilization, and application data. This information can be accessed through the links provided below.

Fluorochrome Data Table

As a guide to fluorophores for confocal and widefield fluorescence microscopy, the table presented in this section lists many commonly-used fluorochromes, with their respective peak absorption and emission wavelengths and suggested laser illumination sources. While the authors assume responsibility for the accurate reporting of the data as published in various reliable sources, several caveats must be given. In reviewing the literature, one will frequently find somewhat different data supplied for the identical fluorochrome.

Contributing Authors

Mortimer Abramowitz - Olympus America, Inc., Two Corporate Center Drive., Melville, New York, 11747.

Christopher Hardee, Roy Kinoshita, Travis Wakefield, and Robert Johnson - Omega Optical, Inc., 210 Main Street, Brattleboro, Vermont, 05301.

Turan Erdogan - Semrock, Inc., 3625 Buffalo Road, Rochester, New York, 14624.

Douglas B. Murphy - Department of Cell Biology and Anatomy and Microscope Facility, Johns Hopkins University School of Medicine, 725 N. Wolfe Street, 107 WBSB, Baltimore, Maryland 21205.

Kenneth R. Spring - Scientific Consultant, Lusby, Maryland, 20657.

Matthew J. Parry-Hill, Thomas J. Fellers, and Michael W. Davidson - National High Magnetic Field Laboratory, 1800 East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310.