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Surface plasma resonance detection device based on laser scanning surface confocal microscopic system

A confocal microscopy, surface plasmon technology, applied in measurement devices, biological testing, material inspection products, etc., can solve problems such as further improvement of limitations, fluorescence bleaching phenomenon, inability to accurately obtain kinetic information, etc., to improve the signal-to-noise ratio. and detection sensitivity, noise reduction effect

Active Publication Date: 2009-05-27
TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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Problems solved by technology

However, the laser scanning confocal system also has its limitations: such as the inability to accurately obtain the kinetic information of the interaction (kinetic constants and equilibrium constants), the existence of fluorescence bleaching, the difficulty of quantitative and sensitive detection, etc.
These limitations limit the further improvement of this system and its application in other fields such as materials science, nanosystems and supramolecular aggregates

Method used

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  • Surface plasma resonance detection device based on laser scanning surface confocal microscopic system

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Embodiment 1

[0035] See figure 1 . Surface plasmon resonance detection device based on laser scanning confocal microscopy system, including sample platform, hemispherical prism, constant temperature flow cell, inverted laser scanning confocal imaging microscope, syringe pump, electronic control angle device, data acquisition and processing system.

[0036] A sample platform with a three-dimensional manual adjustment mechanism. There is a hemispherical prism, the core component of surface plasmon resonance, on the sample platform. Below the hemispherical prism, there is a constant temperature flow cell 5 with a size of 15mm×15mm×5mm and a volume of 1125 (mm) 3 The constant temperature flow cell adopts polytetrafluoroethylene material as the four walls, the bottom surface is made of quartz glass, and the rubber sealing ring is sealed; the liquid inlet of the constant temperature flow cell 5 is connected with the liquid outlet of the syringe pump 7 through a pipeline, and the constant tempera...

Embodiment 2

[0049] The silicon photodiode 9 adopts a photomultiplier tube with a model number of 9863 / 100B, and the others are the same as in Embodiment 1. The sensitivity and detection accuracy of the device in this embodiment can be improved by 2 orders of magnitude.

Embodiment 3

[0051] The gold film plated on the outer surface of the flat bottom of the hemispherical prism is replaced by a silver film, and the surface is chemically modified with antibodies to the core protein p24 of HIV-1, and HIV-1 flows in the constant temperature flow cell The antigen of type (HIV-1) core protein p24, others are identical with embodiment 1, can make diagnosis detection to HIV-1 type.

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Abstract

The invention relates to a surface plasma resonance device for observing and detecting the biomolecular reaction in real time based on laser scanning confocal microscopic system. The device is configured in such a manner that a sample stage is provided with 3-dimensional manually-adjustable mechanism, a hemispherical prism as the surface plasma resonance core part is arranged on the sample stage, a constant-temperature flow through cell of which the pipe is connected with an injection pump is disposed under the hemispherical prism; the sample stage is placed on the position where the objective table of an inverted laser scanning confocal imaging microscope is arranged; an electrically-controlled angle adjusting device comprises a turntable platform controlled by a step motor and mechanical arms on the turntable platform; one mechanical arm is provided with a laser for generating surface plasma resonance, a beam-expanding mirror assembly, and a polarizer, which are arranged sequentially along the light path direction of the exit light, and the other mechanical arm is provided with a focusing lens and a silicon photodiode which are arranged sequentially along the light path direction of the reflected light; the silicon photodiode and a phase-locking amplifier are connected via data line, and the phase-locking amplifier is connected with a data acquisition and processing system which is connected with the controller of the injection pump.

Description

technical field [0001] The invention belongs to a device for detecting biomolecular reactions, in particular to a surface plasmon resonance device for real-time observation and detection of biomolecular reactions based on a laser scanning confocal microscope system. Background technique [0002] In life science research, biomolecular interaction is a basic life phenomenon and one of the major issues in modern life science research. There are many traditional methods for studying interaction, such as radioimmunoassay, enzyme-linked immunoassay, labeling tracer, etc. However, because these processes involve different types and contents of cells and biomolecules, and there are complex interactions between various substances, it is difficult to accurately obtain relevant transmission information using these traditional research methods. At the same time, increasing new Protein and DNA sequence data also urgently require methods for accurate and rapid identification of biologica...

Claims

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Application Information

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IPC IPC(8): G01N21/55G01N21/45G01N21/64G01N33/48G01N21/552
Inventor 张洪艳汪鹏飞吴世康
Owner TECHNICAL INST OF PHYSICS & CHEMISTRY - CHINESE ACAD OF SCI
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