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Fluorescence-related spectrum measurement system

A spectral measurement and fluorescence-related technology, which is applied in the direction of fluorescence/phosphorescence, measuring devices, and material analysis through optical means, can solve the problems of inconvenient measurement of fluorescence-related spectra, reduce measurement costs, reduce processing requirements, and eliminate space problems. The effect of the restriction

Inactive Publication Date: 2019-03-26
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Due to the limitations of these factors, it brings a lot of inconvenience to the measurement of fluorescence correlation spectrum.

Method used

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  • Fluorescence-related spectrum measurement system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Such as figure 1 As shown, a fluorescence correlation spectroscopy measurement system includes an excitation light source 7, a first lens 6, a first color filter 5, a dichroic mirror 4, an objective lens 3, an optical fiber 2, an optical fiber cone 1, and a second color filter 8, the second lens 9 and the detector 10; the end face of the fiber cone 1 is provided with a nanohole 11; the laser light emitted by the excitation light source 7 passes through the first lens 6, the first color filter 5 and the dichroic mirror 4 enters the objective lens 3, then focuses into the optical fiber 2 and the fiber cone 1 through the objective lens 3, and finally forms a spatially confined excitation in the nanohole 11; the excited fluorescence in the nanohole 11 passes through the optical fiber 2, the objective lens 3, and the dichroic mirror 4 , the second color filter 8 and the second lens 9 enter the detector 10 .

[0027] The fluorescence correlation spectroscopy measurement syst...

Embodiment 2

[0039] Fluorescence correlation spectrum measurement and calculation of fluorescent dye solution:

[0040] use figure 1 The device shown is used to measure the fluorescence correlation spectrum of the fluorescent dye FITC solution by time-correlated single photon counting method.

[0041] The 488nm laser transmits the laser light to the tip of the fiber cone through the optical fiber. The volume of the effective detection area of ​​the fiber cone tip is the volume of the excitation light and the volume of the receiving cone, which can reach the femtoliter range. The fluorescence generated by the excited FITC sample molecules is transmitted through the fiber. To the single photon counting system, the single photon counting system detects the number of photons within a period of time, discretizes it into a digital signal and stores it in the memory, and the computer reads the information in the memory, and calculates the fluorescence autocorrelation function according to the tim...

Embodiment 3

[0047] Combined application of fluorescence correlation spectroscopy and microfluidic chips:

[0048] use figure 1 The shown device measures the fluorescence correlation spectrum of a 10 mmol / L fluorescent dye FITC solution in a microfluidic channel. Insert the fiber tip of the taper into the channel of the microfluidic chip, make the nanohole at the fiber tip contact with the FITC solution, and use the 488nm excitation light to excite the fluorescent dye in the nanohole through the fiber.

[0049] Put the same concentration of FITC solution into the centrifuge tube, and use the optical fiber to go deep into the solution to detect the fluorescence correlation spectrum. Comparing the normalized curves obtained by the two systems and performing fitting can analyze the basic differences in the structural parameters and diffusion time of the two systems, so as to analyze the reflection effect of the microfluidic chip pipe wall on the fluorescence or the effect of the channel size...

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Abstract

The invention belongs to the technical field of spectrum detection systems and in particular relates to a fluorescence-related spectrum measurement system. Exciting light is limited within a flying volume by utilizing a tapered optical fiber, and meanwhile, a fluorescence signal is collected by utilizing an optical fiber, so that time-related photo counting is performed, and furthermore, a fluorescence-related spectrum is acquired. By using the method, a fluorescence confocal microscope, a total internal reflection fluorescence microscope or a two-photon fluorescence microscope is prevented from being used, so that the measurement cost of a fluorescence-related spectrum is reduced; meanwhile, the optical fiber can be penetrated into a solution, a reactor, a microfluidic chip, a cell or a body, so that space imitation of imaging equipment is eliminated; and the fluorescence-related spectrum measurement system can be widely applied to the aspects such as reaction dynamic research, livingcell analysis, biological detection, disease diagnosis and drug screening in the fields of life sciences, physics and chemicals.

Description

technical field [0001] The invention belongs to the technical field of spectrum detection systems, in particular to a fluorescence correlation spectrum measurement system. Background technique [0002] With the development of life science and optical measurement technology, people have begun to study the interaction between substances and the life process at the molecular level. One approach to single-molecule detection is the use of fluorescence correlation spectroscopy. Single-molecule detection using fluorescence correlation spectroscopy can obtain information such as diffusion constants and chemical kinetic parameters, and can also be used in life sciences, live cell analysis, biological detection, disease diagnosis, and drug screening. [0003] Fluorescence correlation spectroscopy (FCS) technology is a kind of fluorescence spectroscopy technology that uses the fluctuation of fluorescence intensity with time to analyze and detect. Fluorescence correlation spectroscopy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64
CPCG01N21/6402G01N21/645G01N2021/6484
Inventor 赵祥伟李绍华崔玉军
Owner SOUTHEAST UNIV
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