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Influence of Fluorochrome Labeling Density on the Photobleaching Kinetics of Fluorescein in Microscopy by Loling Song; R. P. M. van Gijlswijk; I. Ted Young; Hans J. Tanke is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

What is Influence of Fluorochrome Labeling Density on the Photobleaching Kinetics of Fluorescein in Microscopy about?

The objective of this study was to identify, through kinetic analysis of individual elementary reactions, the conditions under which a simple first-order photobleaching kinetic model is sufficient for quantitative fluorescence measurements, and those under which more complex photobleaching kinetics must be considered. Three model systems of various fluorophore densities and distributions were employed to verify the kinetic analysis. The results showed that the photobleaching kinetics of free fluorescein at concentrations lower than 5 mM corresponded closely to a single exponential function and therefore involved predominantly simple unimolecular or pseudounimolecular photochemical reactions. When fluorescein was bound to polyvinyl alcohol (PVA) molecules, the photobleach-ing kinetics of the densely labeled PVA deviated more from a single-exponential function than sparsely labeled PVA. When fluorescein was bound to a DNA probe, the photobleaching kinetics were more complex and deviated significantly from a single-exponential function, due to one or more bimolecular processes with apparent concentrationdependent photobleaching rate constants. The practical applications of time-integr

Who reads Influence of Fluorochrome Labeling Density on the Photobleaching Kinetics of Fluorescein in Microscopy?

Author

Loling Song; R. P. M. van Gijlswijk; I. Ted Young; Hans J. Tanke

Publisher

John Wiley and Sons; Wiley (John Wiley & Sons); John Wiley & Sons Inc.; John Wiley & Sons Ltd.; Wiley (ISSN 0196-4763)

Published

1997

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)