Method and device for improving definition of single-molecule fluorescence imaging

Abstract

The invention relates to a fluorescence spectrum imaging technique, aiming at solving the problem of fuzzy single-molecule fluorescence imaging arising from the fluctuation in molecular fluorescent radiation and lower signal-to-noise ratio. The invention provides a method and device for improving the definition of single-molecule fluorescence imaging. The method comprises the following steps of: scanning and irradiating a single-molecule sample by using laser which is subjected to intensity modulation by an acousto-optic modulator; receiving fluorescent photons emitted by the single-molecule sample and converting the fluorescent photons into a standard logic electrical pulse output signal by using a single-photon detector; inputting the logic electrical pulse output signal to a lock-in amplifier, meanwhile, inputting a synchronous sine waveform modulated voltage signal to the lock-in amplifier; and demodulating the logic electrical pulse output signal to obtain a detected fluorescence spectrum signal and imaging the fluorescence spectrum signal. According to the invention, the intensity modulation to the single-molecule fluorescent photons is achieved by using an intensity modulated excitation light field, the quantum fluctuation in photon counting is eliminated by using analog signals demodulated by the lock-in amplifier, the signal-to-noise ratio is increased, and therefore, the high-definition single-molecule fluorescence imaging is achieved.

Description

technical field [0001] The invention relates to fluorescence spectrum imaging technology, in particular to a method and device for improving the definition of single-molecule fluorescence imaging. Background technique [0002] The formation and development of single-molecule science in the 1990s enabled people to observe and manipulate the microscopic material world at the atomic and molecular scales. Optical measurements at the single-molecule level can reveal individual molecular activity properties that are averagely masked by large molecular ensembles. Single-molecule high-resolution fluorescence imaging for molecular probes and fluorescent dyes is an important means and technology for studying molecular diffusion, gene recombination, protein dynamics and other issues. [0003] Single-molecule fluorescence imaging is generally measured by optical confocal microscopy experimental system. However, due to the radiation fluctuation of single-molecule fluorescence (depending...

AI Technical Summary

Problems solved by technology

[0004] The invention provides a method and method for improving the definition of single-molecule fluorescence imaging, which can solve the problem of blurred single-molecule fluorescence imaging caused by fluctuations in molecular fluorescence radiation and low signal-to-noise ratio

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