| Office | 089/2180-77547 (Frau Steger) |
| Telephone | 089/2180-77537 |
| Room | E1.003 |
| Patrick.Schueler “at” cup.uni-muenchen.de |
Publications
Tim Schröder; Johann Bohlen; Sarah E Ochmann; Patrick Schüler; Stefan Krause; Don C Lamb; Philip Tinnefeld
Shrinking gate fluorescence correlation spectroscopy yields equilibrium constants and separates photophysics from structural dynamics Journal Article
In: Proc Natl Acad Sci U S A, vol. 120, no. 4, pp. e2211896120, 2023, ISSN: 1091-6490.
@article{pmid36652471,
title = {Shrinking gate fluorescence correlation spectroscopy yields equilibrium constants and separates photophysics from structural dynamics},
author = {Tim Schröder and Johann Bohlen and Sarah E Ochmann and Patrick Schüler and Stefan Krause and Don C Lamb and Philip Tinnefeld},
doi = {10.1073/pnas.2211896120},
issn = {1091-6490},
year = {2023},
date = {2023-01-01},
urldate = {2023-01-01},
journal = {Proc Natl Acad Sci U S A},
volume = {120},
number = {4},
pages = {e2211896120},
abstract = {Fluorescence correlation spectroscopy is a versatile tool for studying fast conformational changes of biomolecules especially when combined with Förster resonance energy transfer (FRET). Despite the many methods available for identifying structural dynamics in FRET experiments, the determination of the forward and backward transition rate constants and thereby also the equilibrium constant is difficult when two intensity levels are involved. Here, we combine intensity correlation analysis with fluorescence lifetime information by including only a subset of photons in the autocorrelation analysis based on their arrival time with respect to the excitation pulse (microtime). By fitting the correlation amplitude as a function of microtime gate, the transition rate constants from two fluorescence-intensity level systems and the corresponding equilibrium constants are obtained. This shrinking-gate fluorescence correlation spectroscopy (sg-FCS) approach is demonstrated using simulations and with a DNA origami-based model system in experiments on immobilized and freely diffusing molecules. We further show that sg-FCS can distinguish photophysics from dynamic intensity changes even if a dark quencher, in this case graphene, is involved. Finally, we unravel the mechanism of a FRET-based membrane charge sensor indicating the broad potential of the method. With sg-FCS, we present an algorithm that does not require prior knowledge and is therefore easily implemented when an autocorrelation analysis is carried out on time-correlated single-photon data.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Fluorescence correlation spectroscopy is a versatile tool for studying fast conformational changes of biomolecules especially when combined with Förster resonance energy transfer (FRET). Despite the many methods available for identifying structural dynamics in FRET experiments, the determination of the forward and backward transition rate constants and thereby also the equilibrium constant is difficult when two intensity levels are involved. Here, we combine intensity correlation analysis with fluorescence lifetime information by including only a subset of photons in the autocorrelation analysis based on their arrival time with respect to the excitation pulse (microtime). By fitting the correlation amplitude as a function of microtime gate, the transition rate constants from two fluorescence-intensity level systems and the corresponding equilibrium constants are obtained. This shrinking-gate fluorescence correlation spectroscopy (sg-FCS) approach is demonstrated using simulations and with a DNA origami-based model system in experiments on immobilized and freely diffusing molecules. We further show that sg-FCS can distinguish photophysics from dynamic intensity changes even if a dark quencher, in this case graphene, is involved. Finally, we unravel the mechanism of a FRET-based membrane charge sensor indicating the broad potential of the method. With sg-FCS, we present an algorithm that does not require prior knowledge and is therefore easily implemented when an autocorrelation analysis is carried out on time-correlated single-photon data.
Izabela Kamińska; Johann Bohlen; Renukka Yaadav; Patrick Schüler; Mario Raab; Tim Schröder; Jonas Zähringer; Karolina Zielonka; Stefan Krause; Philip Tinnefeld
Graphene Energy Transfer for Single-Molecule Biophysics, Biosensing, and Super-Resolution Microscopy Journal Article
In: Advanced Materials, vol. 33, no. 24, pp. 2101099, 2021.
@article{Kaminska2021,
title = {Graphene Energy Transfer for Single-Molecule Biophysics, Biosensing, and Super-Resolution Microscopy},
author = {Izabela Kamińska and Johann Bohlen and Renukka Yaadav and Patrick Schüler and Mario Raab and Tim Schröder and Jonas Zähringer and Karolina Zielonka and Stefan Krause and Philip Tinnefeld},
doi = {10.1002/adma.202101099},
year = {2021},
date = {2021-05-01},
urldate = {2021-05-01},
journal = {Advanced Materials},
volume = {33},
number = {24},
pages = {2101099},
publisher = {Wiley},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Stefan Krause; Evelyn Ploetz; Johann Bohlen; Patrick Schüler; Renukka Yaadav; Florian Selbach; Florian Steiner; Izabela Kamińska; Philip Tinnefeld
Graphene-on-Glass Preparation and Cleaning Methods Characterized by Single-Molecule DNA Origami Fluorescent Probes and Raman Spectroscopy Journal Article
In: ACS Nano, vol. 15, no. 4, pp. 6430–6438, 2021.
@article{Krause2021,
title = {Graphene-on-Glass Preparation and Cleaning Methods Characterized by Single-Molecule DNA Origami Fluorescent Probes and Raman Spectroscopy},
author = {Stefan Krause and Evelyn Ploetz and Johann Bohlen and Patrick Schüler and Renukka Yaadav and Florian Selbach and Florian Steiner and Izabela Kamińska and Philip Tinnefeld},
doi = {10.1021/acsnano.0c08383},
year = {2021},
date = {2021-04-01},
urldate = {2021-04-01},
journal = {ACS Nano},
volume = {15},
number = {4},
pages = {6430--6438},
publisher = {American Chemical Society (ACS)},
keywords = {},
pubstate = {published},
tppubtype = {article}
}