Reversible photoswitching enables single-molecule fluorescence fluctuation spectroscopy at high molecular concentration.

We demonstrate that photoswitchable markers enable fluorescence fluctuation spectroscopy at high molecular concentration. Reversible photoswitching allows precise control of the density of fluorescing entities, because the equilibrium between the fluorescent ON- and the dark OFF-state can be shifted...

Ausführliche Beschreibung

Bibliographische Detailangaben
Hauptverfasser: Eggeling, C, Hilbert, M, Bock, H, Ringemann, C, Hofmann, M, Stiel, A, Andresen, M, Jakobs, S, Egner, A, Schönle, A, Hell, S
Format: Journal article
Sprache:English
Veröffentlicht: 2007
_version_ 1826276817749671936
author Eggeling, C
Hilbert, M
Bock, H
Ringemann, C
Hofmann, M
Stiel, A
Andresen, M
Jakobs, S
Egner, A
Schönle, A
Hell, S
author_facet Eggeling, C
Hilbert, M
Bock, H
Ringemann, C
Hofmann, M
Stiel, A
Andresen, M
Jakobs, S
Egner, A
Schönle, A
Hell, S
author_sort Eggeling, C
collection OXFORD
description We demonstrate that photoswitchable markers enable fluorescence fluctuation spectroscopy at high molecular concentration. Reversible photoswitching allows precise control of the density of fluorescing entities, because the equilibrium between the fluorescent ON- and the dark OFF-state can be shifted through optical irradiation at a specific wavelength. Depending on the irradiation intensity, the concentration of the ON-state markers can be up to 1,000 times lower than the actual concentration of the labeled molecular entity. Photoswitching expands the range of single-molecule detection based experiments such as fluorescence fluctuation spectroscopy to large entity concentrations in the micromolar range.
first_indexed 2024-03-06T23:19:33Z
format Journal article
id oxford-uuid:6845d7fd-1085-45d6-8bdd-58e7f2566f94
institution University of Oxford
language English
last_indexed 2024-03-06T23:19:33Z
publishDate 2007
record_format dspace
spelling oxford-uuid:6845d7fd-1085-45d6-8bdd-58e7f2566f942022-03-26T18:43:49ZReversible photoswitching enables single-molecule fluorescence fluctuation spectroscopy at high molecular concentration.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:6845d7fd-1085-45d6-8bdd-58e7f2566f94EnglishSymplectic Elements at Oxford2007Eggeling, CHilbert, MBock, HRingemann, CHofmann, MStiel, AAndresen, MJakobs, SEgner, ASchönle, AHell, SWe demonstrate that photoswitchable markers enable fluorescence fluctuation spectroscopy at high molecular concentration. Reversible photoswitching allows precise control of the density of fluorescing entities, because the equilibrium between the fluorescent ON- and the dark OFF-state can be shifted through optical irradiation at a specific wavelength. Depending on the irradiation intensity, the concentration of the ON-state markers can be up to 1,000 times lower than the actual concentration of the labeled molecular entity. Photoswitching expands the range of single-molecule detection based experiments such as fluorescence fluctuation spectroscopy to large entity concentrations in the micromolar range.
spellingShingle Eggeling, C
Hilbert, M
Bock, H
Ringemann, C
Hofmann, M
Stiel, A
Andresen, M
Jakobs, S
Egner, A
Schönle, A
Hell, S
Reversible photoswitching enables single-molecule fluorescence fluctuation spectroscopy at high molecular concentration.
title Reversible photoswitching enables single-molecule fluorescence fluctuation spectroscopy at high molecular concentration.
title_full Reversible photoswitching enables single-molecule fluorescence fluctuation spectroscopy at high molecular concentration.
title_fullStr Reversible photoswitching enables single-molecule fluorescence fluctuation spectroscopy at high molecular concentration.
title_full_unstemmed Reversible photoswitching enables single-molecule fluorescence fluctuation spectroscopy at high molecular concentration.
title_short Reversible photoswitching enables single-molecule fluorescence fluctuation spectroscopy at high molecular concentration.
title_sort reversible photoswitching enables single molecule fluorescence fluctuation spectroscopy at high molecular concentration
work_keys_str_mv AT eggelingc reversiblephotoswitchingenablessinglemoleculefluorescencefluctuationspectroscopyathighmolecularconcentration
AT hilbertm reversiblephotoswitchingenablessinglemoleculefluorescencefluctuationspectroscopyathighmolecularconcentration
AT bockh reversiblephotoswitchingenablessinglemoleculefluorescencefluctuationspectroscopyathighmolecularconcentration
AT ringemannc reversiblephotoswitchingenablessinglemoleculefluorescencefluctuationspectroscopyathighmolecularconcentration
AT hofmannm reversiblephotoswitchingenablessinglemoleculefluorescencefluctuationspectroscopyathighmolecularconcentration
AT stiela reversiblephotoswitchingenablessinglemoleculefluorescencefluctuationspectroscopyathighmolecularconcentration
AT andresenm reversiblephotoswitchingenablessinglemoleculefluorescencefluctuationspectroscopyathighmolecularconcentration
AT jakobss reversiblephotoswitchingenablessinglemoleculefluorescencefluctuationspectroscopyathighmolecularconcentration
AT egnera reversiblephotoswitchingenablessinglemoleculefluorescencefluctuationspectroscopyathighmolecularconcentration
AT schonlea reversiblephotoswitchingenablessinglemoleculefluorescencefluctuationspectroscopyathighmolecularconcentration
AT hells reversiblephotoswitchingenablessinglemoleculefluorescencefluctuationspectroscopyathighmolecularconcentration