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System for detecting microbial interface adhesion based on surface plasmon imaging and method

A surface plasmon, detection system technology, applied in measurement devices, material analysis by optical means, instruments, etc., can solve the problems of signal interference, inability to truly reflect the bacterial adhesion process, and lack of spatial resolution.

Inactive Publication Date: 2019-04-16
UNIV OF SCI & TECH OF CHINA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the above methods still have many deficiencies in the field of microbial interfacial adhesion.
QCM replaces various biological indicators of the sample with the overall average value, and lacks spatial resolution. This technology ignores key dynamic processes and the heterogeneity of biological samples, and cannot perform real-time dynamic observation of single cells in the study of microbial interface adhesion.
TIRFM needs to label bacteria with fluorescent proteins, and fluorescent proteins may affect the metabolic activity of bacteria, which cannot truly reflect the adhesion process of bacteria from the planktonic state to the surface state. At the same time, the signal of the sample is easily interfered by background fluorescent molecules, resulting in Data with low signal-to-noise ratio

Method used

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  • System for detecting microbial interface adhesion based on surface plasmon imaging and method
  • System for detecting microbial interface adhesion based on surface plasmon imaging and method
  • System for detecting microbial interface adhesion based on surface plasmon imaging and method

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Embodiment 1

[0035] Example 1. A system for detecting microbial interface adhesion based on surface plasmon imaging

[0036] Such as figure 1As shown, the detection system includes a surface plasmon resonance microscopic imaging system provided with a plasmon resonance sensor chip and a self-assembled monolayer, a polydimethylsiloxane reaction chip set on the plasmon resonance sensor chip pool.

[0037] Wherein the surface plasmon resonance microscopic imaging system includes a laser generator, an incident angle adjustment component, an optical microscopic magnifying objective lens, a plasmon resonance sensor chip, and an image sensor arranged sequentially along the optical path.

[0038] The plasmon resonance sensor chip is composed of a base material, a chromium layer, and a gold layer arranged in sequence. The specific preparation method is to first coat the upper surface of the glass substrate with a 2nm chromium layer, and then coat the upper surface of the chromium layer with a 48nm...

Embodiment 2

[0043] Embodiment 2, the detection of the adhesion kinetics of escherichia coli

[0044] Step 1, inoculate a single colony of Escherichia coli on the solid medium into 50 mL of LB medium, and cultivate for 12 hours to reach the plateau stage. Then the LB bacterial liquid was centrifuged at 6000 g for 5 min, and the supernatant was filtered off. Then add 5.7mM potassium chloride solution to the centrifuge tube to resuspend the bacteria, centrifuge the bacteria solution at the same speed and time, filter the supernatant again, repeat the above operation two to three times to wash away the LB medium. Finally, continue to add 5.7mM potassium chloride solution until the bacterial solution OD600=0.2, and set aside.

[0045] Step 2: Add 500 μL of the 5.7mM potassium chloride bacterial solution described in step 1 to the PDMS reaction cell on the plasmon resonance sensor chip, turn on the laser generator, SPR microscope, and image sensor respectively, adjust the laser intensity to 11...

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Abstract

The invention belongs to the field of environmental science and engineering, and discloses a system for detecting microbial interface adhesion based on surface plasmon imaging and method. The system for detecting microbial interface adhesion based on surface plasmon imaging is based on surface plasmon resonance microscopy (SPRM) and self-assembled monolayers (SAMs), advantages of being label-free,real-time in observation and high in time resolution are achieved, the adhesion kinetic parameters of bacteria on different interfaces such as an adhesion speed and a kinematic phenotype can be accurately determined by an online counting method, and the time constant of adhesion to the surface can be obtained by calculating and fitting.

Description

technical field [0001] The invention belongs to the field of environmental science and engineering, in particular to a system and method for detecting microbial interface adhesion based on surface plasma imaging. Background technique [0002] Biofilm is an important form of microbial aggregate growth. In the process of wastewater treatment, on the one hand, it is necessary to develop a carrier with good biocompatibility, which is conducive to the formation of biofilm and improve the efficiency of wastewater treatment; on the other hand, in the membrane bioreactor, it is necessary to control the formation of biofilm on the membrane surface to prevent Irreversible fouling of the membrane. Therefore, effective regulation of biofilm is of great significance for water treatment process. It is generally believed that the initial adhesion of microorganisms on solid interfaces affects the formation of biofilms, and understanding the adhesion process of microorganisms on interfaces...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/552
CPCG01N21/553
Inventor 刘贤伟张婷刘轶男
Owner UNIV OF SCI & TECH OF CHINA
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