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...
Hauptverfasser: | , , , , , , , , , , |
---|---|
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 |