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

A fluorescence imaging and single-molecule technology, which is applied in the field of fluorescence spectral imaging, can solve problems such as low signal-to-noise ratio, molecular fluorescence radiation fluctuations, single-molecule fluorescence imaging blur, etc., to improve signal-to-noise ratio, eliminate quantum fluctuations, and achieve high-definition degree of effect

Inactive Publication Date: 2012-06-20
SHANXI UNIV
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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 pr

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

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[0015] The method and device of the present invention will be further described in detail below by taking SR dye molecule samples as an example with reference to the accompanying drawings.

[0016] Sample preparation: firstly, the concentration of the polymer PMMA should be prepared according to the thickness of the film required by the experiment. Put the cleaned coverslip on the tray of the glue homogenizer, turn on the power of the glue homogenizer, turn on the vacuum pump to adsorb the slides on the tray, drop the prepared toluene solution containing PMMA and SR dyes onto the coverslip, Turn on the start button of the homogenizer The homogenizer will rotate at the number of revolutions set in advance. According to the thickness of the sample film, the speed is usually set at 800-3000 revolutions per minute, and the rotation time needs to be adjusted according to the concentration of the sample ( eg ~1 min). After labeling the sample glass slide coated with polymer, put it...

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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...

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Application Information

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IPC IPC(8): G01N21/64
Inventor 张国峰陈瑞云高岩肖连团贾锁堂
Owner SHANXI UNIV
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