Fluorescence single-molecule chemistry

Fluorescence microscopy techniques are now available which are capable of studying supramolecular interfacial assemblies with excellent spatial and temporal resolution as well as exceptional sensitivity. These methods were initially developed for use in cellular biology, but are finding increasing application in interfacial supramolecular chemistry. This trend is set to continue as methods in single-molecule spectroscopy and time-resolved microscopy evolve. 

Lakowicz, J. and Fu, Y. (2006) Enhanced Fluorescence of Cy5-Labeled DNA Tethered to Silver Island Films Fluorescence Images and Time-Resolved Studies Using Single-Molecule Spectroscopy Analytical Chemistry 78 6238 - 6245. 

Nie, S.M., Chiu, D.T., and Zare, R.N. (1995) Real-time detection of single-molecules in solution by confocal fluorescence microscopy. Analytical Chemistry, 61, 2849-2857. 

Wu, A.H.B., Fukushima, N., Puskas, R., Todd, J., and Goix, P. (2006) Development and preliminary clinical validation of a high sensitivity assay for cardiac troponin using a capillary flow (single molecule) fluorescence detector. Clinical Chemistry, 52, 2157 2159. 

Recent advances in ultrasensitive instrumentation have allowed the detection of individual atoms and molecules in solids [174, 175], on surfaces [176, 177], and in the condensed phase [178, 179] using laser-induced fluorescence. In particular, single molecule detection in the condensed phase enables scientists to explore new frontiers in many scientific disciplines, such as chemistry, molecular biology, molecular medicine and nanostructure materials. There are several optical methods to study single molecules, the principles and application of which have been reviewed by Nie and Zare [180]. These methods are listed in Tab. 6.12. 

Dittrich, P S. and Manz, A., Single-molecule fluorescence detection in microfluidic channels— The Holy Grail in xTAS7, Anal Bioanalytical Chemistry, 382, 1771-1782, 2005. 

Single molecule studies. All the techniques discussed so far provide knowledge about the behaviour of nanocrystals at an ensemble level. Unlike molecules that are identical to each other, several key parameters such as surface chemistry can vary from nanocrystal to nanocrystal. This makes it essential to supplement ensemble information with other types of spectroscopic tools that can convey information regarding speciflc members. Due to the relatively small absorption cross sections of individual members, fluorescence based optical techniques have been frequently used to probe individual members or populations. 

This book is aimed at experimental scientists with a physical chemistry or biochemistry background who wish to enter this new and exciting field of research and to apply single molecule fluorescence techniques to studies of macromolecular structure and function. The book is designed to present an introduction to the topic, from the practical implementation of single molecule fluorescence experiments, through methods of data analysis to a description of a range of current and future... 

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