Fluorescence detection of single molecules applicable to small volume assays

Enderlein, Jörg; Ambrose, W. P.; Goodwin, Peter M.; Keller, Richard A.

doi:10.1007/978-3-0348-8817-2_13

Cited by 5 publications

(2 citation statements)

References 153 publications

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“…Detection and spectroscopy of single molecules at both cryogenic 1,2 and ambient [3][4][5][6][7][8][9][10][11][12] temperatures has attracted considerable attention in the past few years. First appearing as a branch of fundamental spectroscopy physics, the field has evolved rapidly toward various applications both in the biological sciences and in applied physics.…”

Section: Introductionmentioning

confidence: 99%

Fluorescence correlation spectroscopy on a mirror

Rigneault

Lenne

2003

J. Opt. Soc. Am. B

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Fluorescence correlation spectroscopy when fluorescent nanoemitters are diffusing in the vicinity of a dielectric mirror is studied both theoretically and experimentally. We demonstrate that two important effects occur when the confocal detection volume is located on the mirror's surface. First, the count rate per emitter is significantly enhanced owing to control of spontaneous emission and enhancement of the excitation field. Second, interference fringes in the excitation beam give rise to a new characteristic time in the photocurrent's autocorrelation function. This new time is found to be independent of the transverse excitation field's beam waist and permits accurate measurement of diffusion coefficients without any a priori knowledge of the confocal volume geometry.

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Section: Introductionmentioning

confidence: 99%

Fluorescence correlation spectroscopy on a mirror

Rigneault

Lenne

2003

J. Opt. Soc. Am. B

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“…In recent years, the fluorescence detection of single molecules (SMD) under ambient conditions within a liquid environment has seen a tremendous development, see e.g. [1][2][3][4][5]. After an initial period of establishing the techniques of single molecule fluorescence detection, the main interest has increasingly shifted towards the application of this method to studying biologically interesting problems, such as enzyme kinetics and single DNA conformational dynamics (for some recent papers see e.g.…”

Section: Introductionmentioning

confidence: 99%

Positional and Temporal Accuracy of Single Molecule Tracking

Enderlein

2000

Single Mol.

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In a recent paper, a new method for tracking fluorescing single molecules that diffuse within bilipid layers or cell membranes was presented and its efficiency theoretically studied. In the present paper, two questions concerning this method are addressed: First, the fundamental question of how accurately can the spatial position of the tracked molecule be determined? Second, is it possible to extract temporal information about e.g. the reaction dynamics of a single fluorescence-labeled enzyme from the signal that is measured during single molecule tracking? The answers to these questions are important when estimating the usefulness of the proposed tracking method as an alternative to conventional single molecule tracking with wide field fluorescence microscopes.

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Fluorescence Spectroscopy of Single Molecules under Ambient Conditions: Methodology and Technology

Böhmer

Enderlein

2003

ChemPhysChem

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This review presents an overview of the fluorescence detection and spectroscopy of single molecules (SMS) in liquids and on surfaces under ambient conditions. The various techniques of SMS, such as confocal epifluorescence detection and wide-field imaging are presented and discussed, together with the different methods of data analysis such as fluorescence correlation spectroscopy and burst-by-burst analysis. Selected applications of the various techniques in physics, chemistry, and biology are described.

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Fluorescence detection of single molecules applicable to small volume assays

Cited by 5 publications

References 153 publications

Fluorescence correlation spectroscopy on a mirror

Fluorescence correlation spectroscopy on a mirror

Positional and Temporal Accuracy of Single Molecule Tracking

Fluorescence Spectroscopy of Single Molecules under Ambient Conditions: Methodology and Technology

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