LRSPR-fluorescence imaging parallel detection device and LRSPR chip manufacturing method

Abstract

The invention provides an LRSPR-fluorescence imaging parallel detection device, which comprises an LRSPR optical device, a fluorescence imaging device, a mechanical device, a data collection device and a computer control device, wherein the LRSPR optical device or the fluorescence imaging device is used for generating resonance and exciting a sample to be tested to emit fluorescence signals; the fluorescence imaging device is used for detecting the fluorescence signals of the sample to be tested; the mechanical device is used for supporting and regulating the LRSPR optical device and the fluorescence imaging device; the data collection device is used for collecting operation data of the LRSPR optical device, the fluorescence imaging device and the mechanical device; and the computer control device is used for processing the operation data collected by the data collection device. The LRSPR is combined with a fluorescence imaging technology; the fluorescence detection signals can be enhanced by using the evanescent field of the LRSPR; the reaction sites of change in molecules or cells can be specifically found; and more comprehensive quantitative information and kinetic reaction process can be provided for biochemical reaction.

Description

technical field [0001] The invention relates to the technical field of fluorescence detection, in particular to an LRSPR-fluorescence imaging parallel detection device and a method for manufacturing an LRSPR chip. Background technique [0002] Surface Plasmon Resonance (SPR) technology is an advanced and general-purpose label-free biochemical detection technology developed in recent years. The probe detects changes in the optical parameters of the sensing medium. The traditional SPR method to detect biological samples can only detect substances within the range of 200nm of the sensor chip, which is only suitable for the study of surface changes, and cannot detect the internal changes of micron-sized cells. The newer long-range surface plasmon (LRSP) technology has a deeper penetration depth, which can reach the order of several microns, and has a wider selection of samples to be tested, and can be used to detect macromolecules, such as for the detection of proteins It is a...

AI Technical Summary

Problems solved by technology

In recent years, the long-range surface plasmon resonance (LRSPR) in the newer technology has higher sensitivity, longer surface propagation length, higher electric field strength, and sharper angular resonance curve, which wears The penetration depth can reach the order of microns, which is far greater than the penetration depth of the traditional SPR for the analyte. The deeper the penetration, the more sensitive the change of the refractive index of the analysis medium, and the higher the sensitivity. Although it can improve the detection sensitivity, LRSPR as a This label-free detection method depends on the change of the mass of the analyte molecule, and it is difficult to detect small molecules, and what is obtained is a comprehensive phenomenon of the change in the refractive index of the sample to be tested, which limits the further improvement of detection sensitivity.

Fluorescence detection technology is a good specific detection method, but it is difficult to achieve quantitative detection with pure fluorescence detection method. If SPR technology is combined with fluorescence technology, the evanescent field of SPR can be used to enhance the detection signal of fluorescence, and specificity can also be achieved. Finding the reaction site of molecular or cell changes will provide more comprehensive quantitative information and kinetic reaction process for biochemical reactions

The surface plasmon-enhanced fluorescence spectroscopy device based on the converging optical system in the prior art is based on the detection at a fixed angle, and the detection range of the refractive index is limited, which cannot meet the detection of samples with large differences in refractive index. The impact can only detect substances in the range of 200nm, and cannot detect changes in the interior or upper surface of cells at the micron level

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