Homebuilt single-molecule scanning confocal fluorescence microscope studies of single DNA/protein interactions.

Homebuilt single-molecule scanning confocal fluorescence microscope studies of single DNA/protein interactions. Research Abstract Details 

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Homebuilt single-molecule scanning confocal fluorescence microscope studies of single DNA/protein interactions. Abstract Text:

Haocheng Zheng,Lori S Goldner,Sanford H Leuba,

Many technical improvements in fluorescence microscopy over the years have focused on decreasing background and increasing the signal to noise ratio (SNR). The scanning confocal fluorescence microscope (SCFM) represented a major improvement in these efforts. The SCFM acquires signal from a thin layer of a thick sample, rejecting light whose origin is not in the focal plane thereby dramatically decreasing the background signal. A second major innovation was the advent of high quantum-yield, low noise, single-photon counting detectors. The superior background rejection of SCFM combined with low-noise, high-yield detectors makes it possible to detect the fluorescence from single-dye molecules. By labeling a DNA molecule or a DNA/protein complex with a donor/acceptor dye pair, fluorescence resonance energy transfer (FRET) can be used to track conformational changes in the molecule/complex itself, on a single molecule/complex basis. In this methods paper, we describe the core concepts of SCFM in the context of a study that uses FRET to reveal conformational fluctuations in individual Holliday junction DNA molecules and nucleosomal particles. We also discuss data processing methods for SCFM.

Homebuilt single-molecule scanning confocal fluorescence microscope studies of single DNA/protein interactions. Publishing Authors By Initials

H Zheng,LS Goldner,SH Leuba,

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Homebuilt single-molecule scanning confocal fluorescence microscope studies of single DNA/protein interactions. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Methods (San Diego, Calif.)

VOLUME: 41

Page Numbers: 342-52

Journal Abbreviation: Methods

ISSN: 1046-2023

DAY: 3

MONTH: Mar

YEAR: 2007

Homebuilt single-molecule scanning confocal fluorescence microscope studies of single DNA/protein interactions. Information

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LANGUAGE: eng

NlmUniqueID: 9426302

Homebuilt single-molecule scanning confocal fluorescence microscope studies of single DNA/protein interactions. Keywords Mesh Terms:

KEYWORDS: Software

MESH TERMS: chemistry

Chemical & Substance for Abstract: Homebuilt single-molecule scanning confocal fluorescence microscope studies of single DNA/protein interactions. Information

Substance Name: Nucleosomes

Registry Number: 0

Grant and Affiliation Information for Homebuilt single-molecule scanning confocal fluorescence microscope studies of single DNA/protein interactions.

AFFILIATION: Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA.

Country: United States

AGENCY: United States NIGMS

GRANT: R01GM077872

ACRONYM: GM

MEDLINETA: Methods

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