1.Fluorescence in
Biology, Biochemistry and Biophysics
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Fluorescence is a non-destructive, non-invasive,
intrinsic, and highly sensitive tool for monitoring various processes in
biology and medicine.
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Stimulation of acceptor fluorescence and quenching
of donor fluorescence is used to monitor excitation energy migration and
transfer.
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Fluorescence is a highly sensitive, selective,
and rapid analytical probe for measuring concentrations of very dilute
fluorophores.
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Fluorescence can monitor the microenvironment
of fluorescent molecules. Measurements of fluorescence include: (a) lifetime
(information on the dynamics of biological processes); (b) quantum yield
(information on photochemistry; heat loss); (c) polarization (information
on organization) ; (d) action spectra ( information on excitation energy
transfer); and (e) emission spectra
(nature of fluorophores), etc.
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Fluorescence spectrum is a mirror image of absorption
spectrum, but shifted to longer wavelengths
(the Stokes shift)
References:
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George G. Guilbault (1973) Practical Fluorescence,
Theory, Methods and Techniques. Marcel Dekker, Inc., New York.
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David M. Jameson (1984) Fluorescence Principles,
Methodologies and Applications. In Fluorescein Hapten: An Immunological
Probe. E.W. Voss Jr.(Editor) CRC Press, pp. 23-48.
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Joseph R. Lakowicz (1983) Principles of Fluorescence
Spectroscopy. Plenum Press, New York.
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Govindjee, J. Amesz, and D.C. Fork (editors) (1986)
Light Emission by Plants and Bacteria. Academic Press, New York.
2.Historical
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"Luminescence" was first observed by Monardes
in 1565 from the extract of Lignum Nephritcum, (that had been recommended
for curing kidney ailments). However, Althanius Kircher (1646) was the
first one to discuss, at length, its bichromatic appearance. It was yellow
in transmitted light and blue in reflected light; the blue light must have
been its fluorescence.
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Sir David Brewster ( 1834), a Scottish preacher,
noted the red emission from chlorophyll in 1833. While discussing his concept
of the color of natural bodies, he remarked almost in passing "In making
a strong beam of the sun's light pass through the green fluid, I was surprised
to observe that its colour was a brilliant red, complementary to
the green. By making the ray pass through greater thickness in succession,
it became first orange and then…." The green fluid in Brewster's experiment
was an alcohol extract of Laurel leaves. It must have contained chlorophyll,
the pigment of green leaves, as named by Pelletier and Caventou (1818).
I consider it likely that this was not only the discovery of chlorophyll
fluorescence, but also of the phenomenon of reabsorption of fluorescence
in thick samples.
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The clearest discovery of the phenomenon of fluorescence
was that by Sir John Herschel (1845) in a clear solution of quinine sulfate.
He noted the "celestial" blue color of this solution, but, had unfortunately
called it
epipolic dispersion.. The following year Brewster
(1846) designated it as internal dispersion.. It was left to Sir G.G. Stokes
(see below) to call it fluorescence.
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Professor G.G. Stokes (1852), Professor of Mathematics
at Cambridge University, who is well known for his discovery that emission
bands are shifted to wavelengths longer than the absorption bands (the
Stokes shift), first used the term dispersive reflexion, but quickly added
a footnote: "I confess that I do not like this term. I am almost inclined
to coin a word, and call the appearance fluorescence, from fluor-spar,
as the analogous term opalescence is derived from the name of a mineral."[
Latin fluo=to flow + spar= a rock).]Stokes is the first one to recognize
this phenomenon as light emission. Askenasy (1867) has credited Stokes
also for the discovery of both phycobilin and chlorophyll a fluorescence
in fresh red algae.
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The term "Phosphorescence" dates back to the early
1500s and was so named after the Greek "light bearing". In fact,
the element phosphorus was named from the same Greek word in 1669, since
it was found to produce a bright light in a dark room.
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http://cfs.umbi.umd.edu/jrl/weber.htm
provides information on Professor Gregorio Weber, who pioneered Fluorescence
as a tool of biological structure and function; this note on him was written
by J.R. Lakowicz
References:
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Harvey, E.N. (1957) A History of Luminescence
From the Earliest Times Until 1900. The American Philosophical Society,
Philadelphia, PA.
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Govindjee (1995) Sixty-three Years Since Kautsky:
Chlorophyll a Fluorescence. Aust J Plant Physiol 22: 131-160.