期刊
JOURNAL OF PHYSICAL CHEMISTRY LETTERS
卷 13, 期 21, 页码 4823-4830出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.jpclett.2c00896
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资金
- Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) via project A02 of the Collective research Collaboration [SFB1456]
- Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany's Excellence Strategy [EXC 2067/1390729940]
- European Research Council (ERC) via project smMIET under the European Union's Horizon 2020 research and innovation programme [884488]
- EPSRC
We propose a new method that combines fluorescence correlation spectroscopy (FCS) on the microsecond time scale with fluorescence antibunching measurements on the nanosecond time scale for measuring photophysical rate constants of fluorescent molecules. The method allows for accurate measurement of photophysical rate constants of different dyes through theoretical analysis and experiment.
We present a new method that combines fluorescence correlation spectroscopy (FCS) on the microsecond time scale with fluorescence antibunching measurements on the nanosecond time scale for measuring photophysical rate constants of fluorescent molecules. The antibunching measurements allow us to quantify the average excitation rate of fluorescent molecules within the confocal detection volume of the FCS measurement setup. Knowledge of this value allows us then to quantify, in an absolute manner, the intersystem crossing rate and triplet state lifetime from the microsecond temporal decay of the FCS curves. We present a theoretical analysis of the method and estimate the maximum bias caused by the averaging of all quantities (excitation rate and photophysical rates) over the confocal detection volume, and we show that this bias is smaller than 5% in most cases. We apply the method for measuring the photophysical rate constants of the widely used dyes Rhodamine 110 and ATTO 655.
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