Multichannel quantitative detection device and detection method of two-photon fluorescence optical tweezers

A fluorescent optical tweezers, quantitative detection technology, applied in the direction of fluorescence/phosphorescence, material excitation analysis, etc., can solve the problems of high price, limit the popularization and use of instruments, and achieve the effect of improving accuracy

Active Publication Date: 2013-03-27
WUHAN UNIV
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Problems solved by technology

However, the current application of two-photon fluorescence technology is mainly limited to the research of biological tissue imaging by two-photon fluorescence microscopy, and the quantitative detection of important biomolecules and extremely small virus particles (~100 nanometers) in liquid samples has not yet been realized.
In addition, two-photon fluorescence microscopy needs to use expensive femtosecond pulsed lasers as excitation light sources, which also limits the popularization and use of this instrument

Method used

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  • Multichannel quantitative detection device and detection method of two-photon fluorescence optical tweezers
  • Multichannel quantitative detection device and detection method of two-photon fluorescence optical tweezers
  • Multichannel quantitative detection device and detection method of two-photon fluorescence optical tweezers

Examples

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Embodiment 1

[0053] Multi-channel Quantitative Analysis of 5 Influenza A Viruses in the Same Sample

[0054] Using polystyrene microspheres as carriers, the double-antibody immune sandwich method was used to enrich different influenza A viruses respectively. Specifically, the surface of the microspheres was modified with monoclonal antibodies, and the monoclonal antibodies were labeled with fluorescent quantum dots. For details, see image 3 .

[0055] First, immunomicrospheres are prepared. Take the surface carboxyl-modified polystyrene microspheres (16), and couple the H9N2 virus anti-HA monoclonal antibody (17) on its surface by a chemical coupling method to obtain immune microspheres for future use.

[0056] 1) Quantitative detection of H9N2 avian influenza virus

[0057] Enrichment of H9N2 avian influenza virus on immune microspheres: add a certain concentration (10 4 -10 6 pc / mL) of immune microspheres, and added biotinylated monoclonal antibody (18) and quantum dot-streptavidin ...

Embodiment 2

[0063] Combined Detection of 5 Gastric Cancer Markers in the Same Serum Sample

[0064] Polystyrene microspheres enriched with gastric cancer markers were prepared by the same method as in Example 1, and CdSe / ZnS core-shell quantum dots with wavelengths of 545nm, 565nm, 585nm, 605nm and 625nm were used to label the same serum sample respectively Five kinds of gastric cancer markers carcinoembryonic antigen (CEA), gastric cancer antigen MG7-Ag, cytokeratin (CK), vascular endothelial growth factor (VEGF) and thrombin (Thrombin), using embodiment 1 to realize the multiplex of five kinds of viruses Channel Quantitative Detection.

[0065] The enrichment of various gastric cancer marker proteins by microspheres is not limited to the double-antibody immune sandwich method, and various other solutions with good specificity and high capture efficiency such as nucleic acid aptamers can also be used.

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Abstract

The invention provides a multichannel quantitative detection device and a detection method of two-photon fluorescence optical tweezers. The detection device carries out the following steps of: shooting a near-infrared laser beam generated by a near-infrared laser into an objective lens through a beam expander, a beam splitter, a two-tone beam splitter mirror and a dimensional scanning system, and forming the optical tweezers in a sample cell by gathering of the objective lens; shooting a near-infrared scattered light from the optical tweezers into a near-infrared light detector after reflecting the near-infrared scattered light by the two-tone beam splitter mirror and the beam splitter and gathering the near-infrared scattered light by a first focusing mirror; and detecting a fluorescent light penetrating through the two-tone beam splitter mirror after gathering the fluorescent light by a second focusing mirror and splitting the fluorescent light by a beam splitting system. According to the multichannel quantitative detection device, the real-time and quantitative detection can be carried out on metal ions, biological molecules, virus particles and the like, and the simultaneous detection for multiple different to-be-detected objects marked by different quantum dots can be realized; and the miniaturization of the multichannel quantitative detection device is easily realized, and a sample detection method has the advantages of high sensitivity, good selectivity, high speed, little sample dosage, no need of pretreatment and the like.

Description

[0001] technical field [0002] The present invention relates to the field of micro-nano operation technology, the field of optoelectronic technology and the field of biochemical sensing technology, in particular to a two-photon fluorescent optical tweezers multi-channel quantitative detection device and detection method. Background technique [0003] Optical tweezers, that is, single-beam gradient force optical traps, are potential wells formed based on the interaction of scattering force and radiation pressure gradient force, which can capture particles in the entire Mie and Rayleigh scattering range. Gradient force plays a role in trapping particles, and the axial gradient force must overcome the scattering force if the particles are to be stably bound in the optical field potential well. Therefore, it is usually necessary to use a microscope objective lens with a high numerical aperture to highly converge the laser beam, so as to generate a strong enough gradient force t...

Claims

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Application Information

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IPC IPC(8): G01N21/64
Inventor 唐宏武庞代文
Owner WUHAN UNIV
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