Principles of Fluorescence Spectroscopy, Third Edition Joseph R. Lakowicz, 954 pages xxvi, illus., index, ISBN-13: 978-0387-31278-1, Springer, New York 2006 , $89.95, hardcover, with CD-ROM. Reviewed by Barry R. Masters, Visiting Scientist, Department of Biological Engineering, Massachusetts Institute of Technology, Fellow of SPIE and OSA. E-mail: bmasters@ mit.edu Lakowicz has written a full-color, third edition of his classic textbook and reference, Principles of Fluorescence Spectroscopy, hereafter called Principles, that is easily understandable to students, scientists, engineers, and clinicians; speciï¬cally those individuals who have not studied physical chemistry quantum chemistry, molecular spectroscopy, statistical mechanics, and kinetic theory nor organic chemistry, and nevertheless are working with ï¬uorescence techniques and optical microscopes, analytical instruments, diagnostic devices, or are developing new techniques based on these devices. I found the book to be eminently suitable for its intended audience. Lakowicz has extensive experience in ï¬uorescence spectroscopy; he has taught the materials contained in the Principles, and he and his students and colleagues have made many signiï¬cant contributions to the ï¬eld. The well-tested and targeted audience-accepted approach is to teach the principles in a phenomenological manner that combines a concise, yet physically correct, qualitative exposition with carefully designed and executed multicolor diagrams, ï¬gures, and tables that contain experimental data. When necessary, the author provides the required equations that describe the phenomena. Furthermore, the author has designated advanced topics, which may be left out of the syllabus for an introductory course. What is the scope of this book? The content of Principles covers the phenomena that one usually observes in the laboratory with a spectroï¬uorometer, a ï¬uorescence microscope, and a frequency- or time-domain lifetime instrument. The author begins with a description of the components of these instruments: light sources, monochromators, ï¬lters, detectors, and polarizers. The next step is to introduce a variety of ï¬uorophores and then to describe the techniques of characterizing their absorption and emission properties as well as environmental inï¬uences temperature, solvent, quenching on these properties. The amount of material allocated to each topic roughly correlates with the authorâs research interests and the impact of these disparate topics on the work of biological researchers. Therefore, the topics of ï¬uorescence lifetime measurements, anisotropy decays, and resonance energy transfer from small molecules to macromolecules of biological interest predominate. The topics of ï¬uorescence-lifetime microscopy, single-molecule detection, ï¬uorescence correlation spectroscopy, metal-enhanced ï¬uorescence, ï¬uorescence Journal of Biomedical Optics March/April 2008 029901-1 Vol. 13 2 Book Review parts: the ï¬rst part presents the principles of classical ï¬uorescence spectroscopy in which the author presents the phenomena, the theories that explain the phenomena, and the instruments that measure the phenomena; in the second part, the author provides concise perspectives on rapidly emerging topics. The latter topics include the following: multiphoton excition microscopy, ï¬uorescence sensing, new and novel ï¬uorophores, DNA technology, ï¬uorescence-lifetime imaging microscopy FLIM , single-molecule detection, ï¬uorescence correlation spectroscopy FCS , metal-enhanced ï¬uorescence, and surface-plasmon-coupled emission. These latter topics some may call them emerging advanced topics are presented with signiï¬cantly greater theoretical explanations than the material of the ï¬rst part. These latter chapters, although concisely written, are augmented with carefully selected references that should be studied by the reader. The introduction contains historical material on the independent observations made by Herschel, Jablonski, and Stokes. There is a reference to the two German papers published in 1916 by Smoluchowski, and a book that contains the English translations of these papers as well as biographic materials. Many of us use the equations that were ï¬rst derived by Smoluchowski on molecular diffusion and Brownian motion , but we know very little of his scientiï¬c accomplishments during his short life. His biography also includes sideby-side German and English versions of his key papers on diffusion Marian Smoluchowski, His Life and Scientiï¬c Work, Polish Scientiï¬c Publishers, Warsaw, 1986 . This portion of the book helps place the development of ï¬uorescence and its role in science in the proper perspective. It is an interesting and informative section and I would have enjoyed reading even more about others who made major pioneering contributions to ï¬uorescence spectroscopy. The Principles addresses many of the difï¬culties that the beginner spectroscopist confronts in the laboratory. Many a beginner is perplexed when the conversion between wavelength and wave number is required. The authorâs brief discussion of how to proceed is helpful. Anyone who has performed ï¬uorescence measurements based on intensities is well aware of the numerous variables that confound the interpretation of their studies. Many of these problems are resolved when lifetime measurements are performed in either the time domain or the frequency domain. Of course, these techniques are associated with new complexities, especially when there are multiple lifetimes and/or multiple molecular conï¬gurations within the sample volume. Often the novice is confused by the terminology of ï¬uorescence spectroscopy; for instance, when the terms ï¬uorescence anisotropy and ï¬uorescence polarization are taught. Perusal of the chapters on ï¬uorescence anisotropy make for a clear understanding of each of these terms and the associated physical techniques and the assumptions that they are based on. Another insightful example is the section on multiphoton excitation ï¬uorescence in which the author demonstrates that the anisotropy can reach a value of 0.5 and therefore exceed the magnitude of the onephoton limit of anisotropy that is 0.4. The latter half of the book is especially exciting due to the exposition of emerging developments in ï¬uorescence spectroscopy. The text appears to incorporate more of the physics that support the phenomena, the quality of the color illustrations are impressive, and the latter part of the book is written in a more engaging style. Overall, Principles has an appropriate combination of clearly written text, well-designed schematic illustrations, and thoughtful examples of experimental data. All of these features make it a highly recommended and preferred textbook for teachers and beginning students in the ï¬eld of ï¬uorescence spectroscopy. Journal of Biomedical Optics 029901-2 March/April 2008 Vol. 13 2
Journal of Biomedical Optics – SPIE
Published: Mar 1, 2008
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