Basics of fluorescence

Learmonth, Robert P. and Kable, Scott H. and Ghiggino, Kenneth P. (2009) Basics of fluorescence. In: Fluorescence applications in biotechnology and life sciences. Wiley-Blackwell Publishing, Inc., Hoboken, NJ, USA, pp. 1-26. ISBN 978-0-470-08370-3


The objective of this chapter is to provide a simple understanding of molecular spectroscopy, with a focus on concepts and techniques relevant to microscopy. The inherently quantum mechanical concepts of phenomena, including energy levels, radiative and nonradiative processes, and quantum yields, will be developed from a nonmathematical framework that is often used by experts in spectroscopy when discussing general principles. This simplified framework is developed into a practical and useful understanding of optical spectroscopy aimed at nonspecialists. This groundwork supports the more detailed discussion of complex techniques in the following chapters.
To begin the chapter we will outline basic features and properties of light and energy states of molecules. We then connect the two by exploring the interaction
of light with molecules, initially for an isolated molecule. We will consider the key properties of absorption and emission and also those properties that are the bane of microscopy—the so called nonradiative mechanisms. After considering the isolated molecule, we will discuss interactions of molecules with their environment, which develops the concepts underpinning photobleaching and quenching, and also the more recent techniques of FRET and FLIM. En route we will examine the excitation and
emission spectra of typical dyes, which can be found in any dye catalogue (see also Jameson et al. [1] for further reading).

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Item Type: Book Chapter (Commonwealth Reporting Category B)
Refereed: Yes
Item Status: Live Archive
Additional Information: Chapter 1. This publication arose from work and activities supported by the Australian Research Council and National Health and Medial Research Council Research Network grant RN460002.
Faculty / Department / School: Historic - Faculty of Sciences - Department of Biological and Physical Sciences
Date Deposited: 17 Dec 2010 06:39
Last Modified: 07 Sep 2016 03:08
Uncontrolled Keywords: fluorescence; light; absorption; emission; non-radiative decay; spectroscopy; microscopy
Fields of Research : 06 Biological Sciences > 0601 Biochemistry and Cell Biology > 060199 Biochemistry and Cell Biology not elsewhere classified
02 Physical Sciences > 0205 Optical Physics > 020501 Classical and Physical Optics
02 Physical Sciences > 0205 Optical Physics > 020503 Nonlinear Optics and Spectroscopy
10 Technology > 1099 Other Technology > 109999 Technology not elsewhere classified
Socio-Economic Objective: E Expanding Knowledge > 97 Expanding Knowledge > 970103 Expanding Knowledge in the Chemical Sciences
E Expanding Knowledge > 97 Expanding Knowledge > 970110 Expanding Knowledge in Technology
E Expanding Knowledge > 97 Expanding Knowledge > 970111 Expanding Knowledge in the Medical and Health Sciences
E Expanding Knowledge > 97 Expanding Knowledge > 970102 Expanding Knowledge in the Physical Sciences

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