Transient fluorescence lifetime measurement method and measurement system based on single photon counting

Abstract

The invention provides a transient fluorescence lifetime measurement method based on single photon counting, which includes the following steps that: 1) a sample is irradiated by pulse light with high recurrence frequency to excite fluorescence; 2) the fluorescence lifetime of the sample is induced to have transient changes according to a certain period; 3) the fluorescence photons of the sample are detected and the corresponding high-speed digital pulse signals which represent the single photon signals are outputted; 4) the macro times of the photons are divided into different time zones, the photons are allocated to different groups according to the macro time and the groups correspond to the time zones of the macro time of the photons; and 5) time-relevant single photon counting is conducted on the photons in each macro time zone according to the high-speed digital pulse signals so as to get a fluorescence decay curve of the fluorescence photons in the macro time zone relative to time. In such way, the fluorescence decay curves in different macro time zones can be obtained. The invention further provides a corresponding transient fluorescence lifetime measurement system. The transient fluorescence lifetime measurement method and system are capable of recording the transient fluorescence decay dynamic curves of the nanosecond time sequence.

Description

technical field [0001] The present invention relates to the technical field of fluorescence lifetime measurement, in particular, the present invention relates to a transient fluorescence lifetime measurement method and a corresponding measurement system Background technique [0002] Measuring the fluorescence lifetime of labeled and unlabeled fluorophores is one of the important means to study the structure, physical and chemical properties of biological systems. For example, by measuring the fluorescence lifetime of labeled and unlabeled fluorophores, information on the status of tissue metabolism, changes in protein or DNA structures, and interactions between molecules can be obtained. Moreover, imaging with high spatial resolution can also be achieved through the measurement of fluorescence lifetime. It can be seen that the measurement of fluorescence lifetime is one of the important technical means in the field of biophotonics. [0003] Time-correlated single photon co...

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Problems solved by technology

[0011] However, due to the limitations of the signal acquisition dead time of the electronic system and the minimum signal storage time, it is difficult to further reduce the acquisition time of each fluorescence decay kinetic curve. Therefore, the existing TCSPC fluorescence It is difficult for the lifetime measurement system to increase the interval of the time series to the resolution of the nanosecond level, that is, it is difficult to measure the ultrafast process of the transient change of the fluorescence lifetime

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