Biosensing detection by quartz crystal shearing vibration amplitude-modulation frequency-modulation throwing method

A quartz crystal and bio-sensing technology, which is applied in the direction of biological testing, measuring devices, material inspection products, etc., can solve problems such as increased resistance

Inactive Publication Date: 2020-10-23
司士辉
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This model ignores the role of the key itself, adding additional resistance

Method used

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  • Biosensing detection by quartz crystal shearing vibration amplitude-modulation frequency-modulation throwing method
  • Biosensing detection by quartz crystal shearing vibration amplitude-modulation frequency-modulation throwing method
  • Biosensing detection by quartz crystal shearing vibration amplitude-modulation frequency-modulation throwing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Lysozyme aptamer was used to measure lysozyme. A bare quartz crystal with a middle diameter of 8 mm and a diameter of 5 mm was fixed at the bottom of a circular detection cell, and a metal excitation electrode was placed 1 mm away from the exposed back of the quartz crystal. Sealed with waterproof glue, the temperature of the entire detection pool is controlled at a constant temperature of 37 degrees. Wash the crystal surface with absolute ethanol, treat with 1 mg / ml thiol-containing lysozyme aptamer PBS solution overnight at 4 °C, then wash off unfixed lysozyme aptamer with PBS solution, and finally wash with 1 mg / ml lysozyme aptamer ml of bovine serum albumin for blocking. Before starting, use a flow syringe pump to pass through the phosphate buffer solution with pH=7.2 above the quartz crystal, and measure the initial resonance frequency once. During detection, flow 10 mg / ml lysozyme PBS solution at a flow rate of 10 μl / min over the crystal surface modifie...

Embodiment 2

[0047] Example 2 Determination of IgG, the cleaned crystal was immersed in 15 ml of 2-mercaptododecanoic acid ethanol solution at room temperature for 4 hours to form a self-assembled monolayer on the gold surface. During this process, sulfhydryl groups react with gold substrates via Au-S bonds. After washing with ethanol and water, the crystals were sequentially treated with 15 mM NHS and 110 mM EDC aqueous solution for 2 h at room temperature. Fix a pre-treated quartz crystal with a diameter of 8mm and a middle electrode diameter of 5mm on the bottom of a circular detection pool. The periphery of the electrode is sealed with a waterproof rubber ring. Two small holes are left on the top to connect the external liquid, and the liquid is injected with a flow syringe pump. The temperature of the detection pool is controlled at a constant temperature of 37 degrees. Inject pH=7.2 phosphate buffer over the quartz crystal at a rate of 10 μl / min to measure the initial resonance freq...

Embodiment 3

[0048] Example 3 The quartz crystal is sealed with two O-rings pressed together by four screws, and fixed up and down with elastic plastic gaskets. A cylindrical groove is left above the detection cell to facilitate the addition of the detection solution. One side of the crystal is sealed and the other side faces the groove. Drop 50μl of different 10mg / ml protein solutions (BSA, lysozyme, IgG, protein A, etc.) onto the QCM crystal modified by the self-assembly layer, set the excitation voltage to 15V, use the program to set the excitation time to 60s, and then stop Excited for 180s, first measure the resonant frequency when it is not excited, generally it is an approximate straight line with a slight downward slope. When the excitation is not recorded, the frequency is recorded after the excitation is stopped. From this, the frequency-time spectrum can be obtained, and the time when the excitation is stopped can be obtained from the figure. Antigens and antibodies will recombi...

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Abstract

The invention provides a quartz crystal microbalance biosensing method based on a mass load reduction method. A biomolecular probe (such as an antibody and a single-stranded DNA) is modified on a quartz crystal, a detection solution (such as pathogenic bacteria, DNA molecules to be detected and the like) is added to the surface of a sensor, a frequency spectrum scanning system is developed by adopting a direct digital synthesis (DDS) technology, and scanning excitation is carried out on a quartz crystal oscillator according to a preset frequency and sine wave amplitude; and a quartz crystal frequency signal acquisition and analysis system is constructed based on a superheterodyne receiving orthogonal phase-sensitive detection technology, and a QCM response signal amplitude-frequency and phase-frequency characteristic curve is drawn. Quartz crystal microbalance biosensing by a mass load reduction method is realized through a QCM response signal spectrum. According to the method, a detection signal is obtained in situ in the detection liquid, and a quartz crystal microbalance is directly adopted for biosensing detection, so that the method has important application value for medicalinstant detection and on-site rapid detection.

Description

technical field [0001] The invention is a novel biosensor, which belongs to the instrument detection in the field of analytical chemistry, relates to the technical field of sensing and detection, and in particular relates to a mass load decrement method and a quartz crystal microbalance biosensing method. The biological monitoring is carried out through the quartz crystal shear vibration, amplitude modulation and frequency modulation, and different excitation voltages are adjusted to make the crystal vibration amplitude different, and then the detection target on the surface is thrown off, so as to realize quantitative detection or qualitative detection. The technology is simple to operate, has the advantages of no need for any labeling of biological samples, short detection time, high detection sensitivity, etc., and can also realize online, real-time continuous detection, and can be widely used in life science research, drug screening, environmental detection, food detection,...

Claims

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Application Information

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IPC IPC(8): G01N5/04G01N33/53
CPCG01N5/04G01N33/5302
Inventor 司士辉张润万富扶梅冯浪霞余建芳
Owner 司士辉
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