Diving For Science 2005 Proceedings Of The American
Diving For Science 2005 Proceedings Of The American Diving For Science 2005 Proceedings Of The American
Diving For Science 2005 Proceedings Of The American Academy Of Underwater Sciences Figure 1. White-light (left) and fluorescence photographs of two corals (above, Roatan, Honduras) and a bristleworm (below, Bonaire). History The first printed record we have found of fluorescence of a marine organism dates to 1927. A Mr. C. E. S. Phillips (Phillips, 1927) was walking along the shore in Torbay, England, and noticed that the anemones in a tidepool seemed to be an especially bright green. He collected several specimens and used a light with a Wood's glass filter (a filter that absorbs visible light and transmits only ultraviolet) to confirm that it fluoresced under ultraviolet light. Phillips suggested that marine biologists add such a light to their repertoire of research equipment, but nothing came of his suggestion. In the 1930's and '40's Siro Kawaguti, a Japanese marine biologist working at the Palao Tropical Biological Station, studied the pigments of corals. He noted that the most common pigment was a fluorescent green. Kawaguti carried out a number of manipulative experiments on the pigments, and authored several scientific papers on the topic (Kawaguti, 1944, 1966, 1969). SCUBA divers rediscovered fluorescence in the 1950's. Luis Marden, a photographer for National Geographic magazine, wrote in 1956 that he noticed red anemones at a depth of 60 feet, where there should have been no red (Marden, 1956). The red color disappeared in flash photographs, and Marden correctly guessed that the effect was due to fluorescence. Conrad Limbaugh and Wheeler North, working at the Scripps Institution of Oceanography, 2
Diving For Science 2005 Proceedings Of The American Academy Of Underwater Sciences investigated several brightly colored anemones from West Coast waters and determined that they were fluorescent (Limbaugh and North, 1956). The biggest catalyst to the early study of fluorescence, though, was the work of Rene Catala, Director of the Noumea Aquarium in New Caledonia. Catala was the first person to systematically look for fluorescence in corals (Catala, 1958, 1959, 1960, 1964). He didn’t do his looking underwater, though. His staff divers collected specimens during the day and he examined and photographed them under ultraviolet light in the Aquarium at night. Catala became enthusiastic about fluorescence and established a Hall of Fluorescent Corals in the Aquarium. In 1959 he shipped fluorescent corals to Europe and mounted an exhibition in Antwerp, Belgium. Aquarium displays of fluorescing corals have been a curiosity ever since. Possibly the first person to take ultraviolet light into the sea was Richard Woodbridge III. Not the warm waters of the Caribbean for him - Woodbridge built his own underwater ultraviolet lights and tested them in the chilly waters of Maine. He wrote articles for Skin Diver magazine (Woodbridge, 1959, 1961), and also published in the scientific literature (Woodbridge, 1959). Captain Jacques-Yves Cousteau mentions the use of ultraviolet light underwater in The Silent World, but the time when he used it is uncertain. Others have dabbled in fluorescence over the years, in both the scientific and popular realms, including Dr. Ken Read at Boston University (Read, 1967; Read et al., 1968), and Paul Zahl and David Doubilet at National Geographic (Zahl, 1963; Doubilet, 1997), So while the idea of looking at underwater fluorescence is not new, the early attention was sporadic at best, presented mostly as a curiosity, and there has only recently been serious interest in the phenomenon in the marine research community. For a more extensive list of references to both the scientific and popular literature, visit http://www.nightsea.com/references.htm. Principles of fluorescence Fluorescence is defined as the absorption of electromagnetic radiation at one wavelength, followed by its re-emission at another. Notice that I didn't say 'ultraviolet light'. It is a common misconception that fluorescence is specifically associated with ultraviolet. Yes, ultraviolet is very often used to stimulate fluorescence, but that doesn't mean that it is always the best choice. For our own research we use blue lights almost exclusively. To understand how best to see fluorescence we need to understand a few of the measures we use to characterize it. The emission spectrum is the distribution of light energy as a function of wavelength for the emitted light, while the excitation spectrum is a graph of the relative ability of different wavelengths of light to stimulate, or excite, the fluorescence. Figure 2A shows the emission spectra for two of the fluorescent proteins commonly found in corals. We often describe spectra such as these by their peak (the wavelength of maximum energy) and their width, defined as the full width at half maximum intensity (abbreviated as FWHM). We can see that the cyan-fluorescing protein with the peak at 486 nm has a much broader emission than 3
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<strong>Diving</strong> <strong>For</strong> <strong>Science</strong> <strong>2005</strong> <strong>Proceedings</strong> <strong>Of</strong> <strong>The</strong> <strong>American</strong> Academy <strong>Of</strong> Underwater <strong>Science</strong>s<br />
Figure 1. White-light (left) and fluorescence photographs of two corals (above, Roatan, Honduras) and a<br />
bristleworm (below, Bonaire).<br />
History<br />
<strong>The</strong> first printed record we have found of fluorescence of a marine organism dates to 1927. A<br />
Mr. C. E. S. Phillips (Phillips, 1927) was walking along the shore in Torbay, England, and<br />
noticed that the anemones in a tidepool seemed to be an especially bright green. He collected<br />
several specimens and used a light with a Wood's glass filter (a filter that absorbs visible<br />
light and transmits only ultraviolet) to confirm that it fluoresced under ultraviolet light.<br />
Phillips suggested that marine biologists add such a light to their repertoire of research<br />
equipment, but nothing came of his suggestion.<br />
In the 1930's and '40's Siro Kawaguti, a Japanese marine biologist working at the Palao<br />
Tropical Biological Station, studied the pigments of corals. He noted that the most common<br />
pigment was a fluorescent green. Kawaguti carried out a number of manipulative<br />
experiments on the pigments, and authored several scientific papers on the topic (Kawaguti,<br />
1944, 1966, 1969).<br />
SCUBA divers rediscovered fluorescence in the 1950's. Luis Marden, a photographer for<br />
National Geographic magazine, wrote in 1956 that he noticed red anemones at a depth of 60<br />
feet, where there should have been no red (Marden, 1956). <strong>The</strong> red color disappeared in flash<br />
photographs, and Marden correctly guessed that the effect was due to fluorescence. Conrad<br />
Limbaugh and Wheeler North, working at the Scripps Institution of Oceanography,<br />
2
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