Red light, green light: probing single molecules using alternating-laser excitation.

Single-molecule fluorescence methods, particularly single-molecule FRET (fluorescence resonance energy transfer), have provided novel insights into the structure, interactions and dynamics of biological systems. ALEX (alternating-laser excitation) spectroscopy is a new method that extends single-mol...

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Main Authors: Santoso, Y, Hwang, L, Le Reste, L, Kapanidis, A
Format: Journal article
Language:English
Published: 2008
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author Santoso, Y
Hwang, L
Le Reste, L
Kapanidis, A
author_facet Santoso, Y
Hwang, L
Le Reste, L
Kapanidis, A
author_sort Santoso, Y
collection OXFORD
description Single-molecule fluorescence methods, particularly single-molecule FRET (fluorescence resonance energy transfer), have provided novel insights into the structure, interactions and dynamics of biological systems. ALEX (alternating-laser excitation) spectroscopy is a new method that extends single-molecule FRET by providing simultaneous information about structure and stoichiometry; this new information allows the detection of interactions in the absence of FRET and extends the dynamic range of distance measurements that are accessible through FRET. In the present article, we discuss combinations of ALEX with confocal microscopy for studying in-solution and in-gel molecules; we also discuss combining ALEX with TIRF (total internal reflection fluorescence) for studying surface-immobilized molecules. We also highlight applications of ALEX to the study of protein-nucleic acid interactions.
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spelling oxford-uuid:abfdba97-e301-4657-9fdc-966ebf8c318a2022-03-27T03:25:39ZRed light, green light: probing single molecules using alternating-laser excitation.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:abfdba97-e301-4657-9fdc-966ebf8c318aEnglishSymplectic Elements at Oxford2008Santoso, YHwang, LLe Reste, LKapanidis, ASingle-molecule fluorescence methods, particularly single-molecule FRET (fluorescence resonance energy transfer), have provided novel insights into the structure, interactions and dynamics of biological systems. ALEX (alternating-laser excitation) spectroscopy is a new method that extends single-molecule FRET by providing simultaneous information about structure and stoichiometry; this new information allows the detection of interactions in the absence of FRET and extends the dynamic range of distance measurements that are accessible through FRET. In the present article, we discuss combinations of ALEX with confocal microscopy for studying in-solution and in-gel molecules; we also discuss combining ALEX with TIRF (total internal reflection fluorescence) for studying surface-immobilized molecules. We also highlight applications of ALEX to the study of protein-nucleic acid interactions.
spellingShingle Santoso, Y
Hwang, L
Le Reste, L
Kapanidis, A
Red light, green light: probing single molecules using alternating-laser excitation.
title Red light, green light: probing single molecules using alternating-laser excitation.
title_full Red light, green light: probing single molecules using alternating-laser excitation.
title_fullStr Red light, green light: probing single molecules using alternating-laser excitation.
title_full_unstemmed Red light, green light: probing single molecules using alternating-laser excitation.
title_short Red light, green light: probing single molecules using alternating-laser excitation.
title_sort red light green light probing single molecules using alternating laser excitation
work_keys_str_mv AT santosoy redlightgreenlightprobingsinglemoleculesusingalternatinglaserexcitation
AT hwangl redlightgreenlightprobingsinglemoleculesusingalternatinglaserexcitation
AT lerestel redlightgreenlightprobingsinglemoleculesusingalternatinglaserexcitation
AT kapanidisa redlightgreenlightprobingsinglemoleculesusingalternatinglaserexcitation