Title
Topics in Fluorescence Spectroscopy, Vol. 5: Nonlinear and TwoPhotonInduced Fluorescence,Used
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Fluorescence spectroscopy continues its advance to more sophisticated methods and applications. As one looks over the previous decades, its appears that the first practical instruments for timeresolved measurements appeared in the 1970's. The instrumentation and analysis methods for timeresolved fluorescence advanced rapidly throughout the 1980's. Since 1990 we have witnessed a rapid migration of the principles of timeresolved fluorescence to cell biology and clinical appli tions. Most recently, we have seen the introduction of multiphoton excitation, pumpprobe and stimulated emission methods for studies of biological mac molecules and for cellular imaging. These advanced topics are the subject of the present volume. Twophoton excitation was first predicted by Maria GoppertMayer in 1931, but was not experimentally observed until 1961. Observation of twophoton excitation required the introduction of lasers which provided adequate photon density for multiphoton absorption. Since the early observations of twophoton excitation in the 1960s, multiphoton spectroscopy has been limited to somewhat exotic applications of chemical physics, where it is used to study the electronic symmetry of small molecules. Placing one's self back in 1980, it would be hard to imagine the use of multiphoton excitation in biophysics or cellular imaging.
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