Basics of fluorescence

Edited book (chapter)


Learmonth, Robert P., Kable, Scott H. and Ghiggino, Kenneth P.. 2009. "Basics of fluorescence." Goldys, Ewa M. (ed.) Fluorescence applications in biotechnology and life sciences. Hoboken, NJ, USA. Wiley-Blackwell. pp. 1-26
Chapter Title

Basics of fluorescence

Book Chapter CategoryEdited book (chapter)
ERA Publisher ID3809
Book TitleFluorescence applications in biotechnology and life sciences
AuthorsLearmonth, Robert P. (Author), Kable, Scott H. (Author) and Ghiggino, Kenneth P. (Author)
EditorsGoldys, Ewa M.
Page Range1-26
Number of Pages26
Year2009
PublisherWiley-Blackwell
Place of PublicationHoboken, NJ, USA
ISBN9780470083703
Web Address (URL)http://media.johnwiley.com.au/product_data/excerpt/00/04700837/0470083700.pdf
Abstract

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).

Keywordsfluorescence; light; absorption; emission; non-radiative decay; spectroscopy; microscopy
ANZSRC Field of Research 2020310199. Biochemistry and cell biology not elsewhere classified
510302. Classical and physical optics
510203. Nonlinear optics and spectroscopy
310699. Industrial biotechnology not elsewhere classified
Public Notes

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.

Byline AffiliationsDepartment of Biological and Physical Sciences
University of Sydney
University of Melbourne
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