Biophysics and the Challenges of Emerging Threats (Hardcover, 2009 ed.)

Single-molecule techniques eliminate ensemble averaging, thus revealing transient or rare species in heterogeneous systems [1-3]. These approaches have been employed to probe myriad biological phenomena, including protein and RNA folding [4-6], enzyme kinetics [7, 8], and even protein biosynthesis [1, 9, 10]. In particular, immobilization-based fluorescence te- niques such as total internal reflection fluorescence microscopy (TIRF-M) have recently allowed for the observation of multiple events on the millis- onds to seconds timescale [11-13]. Single-molecule fluorescence methods are challenged by the instability of single fluorophores. The organic fluorophores commonly employed in single-molecule studies of biological systems display fast photobleaching, intensity fluctuations on the millisecond timescale (blinking), or both. These phenomena limit observation time and complicate the interpretation of fl- rescence fluctuations [14, 15]. Molecular oxygen (O) modulates dye stability. Triplet O efficiently 2 2 quenches dye triplet states responsible for blinking. This results in the for- tion of singlet oxygen [16-18]. Singlet O reacts efficiently with organic dyes, 2 amino acids, and nucleobases [19, 20]. Oxidized dyes are no longer fluor- cent; oxidative damage impairs the folding and function of biomolecules. In the presence of saturating dissolved O , blinking of fluorescent dyes is sup- 2 pressed, but oxidative damage to dyes and biomolecules is rapid. Enzymatic O -scavenging systems are commonly employed to ameliorate dye instability. 2 Small molecules are often employed to suppress blinking at low O levels.

General

Imprint:	Springer
Country of origin:	Netherlands
Series:	NATO Science for Peace and Security Series B: Physics and Biophysics
Release date:	June 2009
First published:	2009
Editors:	Joseph Puglisi
Dimensions:	235 x 155 x 12mm (L x W x T)
Format:	Hardcover
Pages:	179
Edition:	2009 ed.
ISBN-13:	978-90-481-2366-7
Categories:	Books > Science & Mathematics > Physics > Applied physics & special topics > Biophysics Books > Science & Mathematics > Biology, life sciences > Biochemistry > General Books > Professional & Technical > Biochemical engineering > Biotechnology > General
LSN:	90-481-2366-6
Barcode:	9789048123667

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