An in-situ characterization device and characterization method for the interaction between microorganisms and solid surfaces

A solid surface, microorganism technology, applied in the field of microbiology and materials science, and microscopy, can solve the problems of the influence of microbial activity, the inability to observe the light quenching for a long time, and the inability to observe the real-time movement behavior of microbial individuals, so as to achieve convenient construction. , Wide range of application, simple operation effect

Active Publication Date: 2022-04-26
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, both methods require fluorescent protein labeling, so there are problems such as the impact of phototoxicity on microbial activity, and the inability to observe light quenching for a long time. In addition, in the fluorescence mode, microorganisms and surface patterns cannot be observed under the lens at the same time
The sensitivity of quartz crystal microbalance can reach the nanogram level, and it can sensitively detect the adhesion and desorption behavior of microorganisms, but it reflects the overall state and cannot observe the real-time movement behavior of individual microorganisms (Applied&Environmental Microbiology[J ], 2005, 71(5):2705-2712.)

Method used

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  • An in-situ characterization device and characterization method for the interaction between microorganisms and solid surfaces
  • An in-situ characterization device and characterization method for the interaction between microorganisms and solid surfaces
  • An in-situ characterization device and characterization method for the interaction between microorganisms and solid surfaces

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Embodiment 1

[0055] An in-situ characterization device for the interaction between microorganisms and solid surfaces, including a temperature-controlled hottable 1, a microbial culture tank 2, a glass substrate 3, solid surface materials 4, an inverted microscope 7, an image sensor 8 and a host computer 9, and a temperature-controlled hottable 1 It is a cuboid container with an opening at the top, and a temperature-controlling heating platform cover 1-1 is provided at the opening at the top, and a circular through hole is provided on the bottom surface of the inner wall of the temperature-controlling heating platform 1; device, the bottom end is fixed on the bottom surface of the inner wall of the temperature control hot table 1, the top opening is provided with a microbial culture tank cover 2-1, and the lower part of the inner wall of the microbial culture tank 2 is provided with a ring-shaped annular baffle 2-2 , the outer edge of the annular baffle 2-2 is fixed along the inner wall of t...

Embodiment 2

[0060] The in-situ characterization device for the interaction between microorganisms and solid surfaces is the same as in Example 1, except that the solid surface material 4 is replaced by a polyurethane film with an area of ​​1cm×1cm and a thickness of 0.05mm, and the surface pattern is hemispheres with a diameter of 2 μm that are hexagonal close-packed shaped raised. The thickness of the glass substrate 3 is 0.17 mm.

[0061] Methods for in situ characterization of microbial interactions with solid surfaces are as follows:

[0062] Put the Staphylococcus aureus in the TSB culture medium, shake the bacteria at 180rpm and 37℃ for 12h, then dilute to 10 5 CFU / ml, take 2ml and add it to the microbial culture tank 2, adjust to 37°C, turn on the inverted microscope 7, adjust the height of the objective lens of the inverted microscope 7 until it focuses on the solid surface 4, turn on the image sensor 8 and the host computer 9, and the host computer 9 The received digital signal...

Embodiment 3

[0065] The in-situ characterization device for the interaction between microorganisms and solid surfaces is the same as in Example 1, except that the solid surface material 4 is replaced by a polystyrene film, with an area of ​​1cm×1cm and a thickness of about 0.05mm; the surface pattern is hexagonal close-packed with a diameter of 5 μm hemispherical bumps. The thickness of the glass substrate 3 is 0.17 mm.

[0066] Methods for in situ characterization of microbial interactions with solid surfaces are as follows:

[0067] Put the Staphylococcus aureus in the TSB culture medium, shake the bacteria at 180rpm and 37℃ for 12h, then dilute to 10 5 CFU / ml, take 2ml and add it to the microbial culture tank 2, adjust to 37°C, turn on the inverted microscope 7, adjust the height of the objective lens of the inverted microscope 7 until it focuses on the solid surface 4, turn on the image sensor 8 and the host computer 9, and the host computer 9 The received digital signal is processed...

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Abstract

The invention relates to an in-situ characterization device and characterization method for the interaction between microorganisms and solid surfaces, and belongs to the interdisciplinary technical field of microscopy, microbiology and materials science. The in-situ characterization device of the present invention includes a temperature-controlled hot stage, a microbial culture tank, a glass substrate, a solid surface material, an inverted microscope, an image sensor and a host computer. The in-situ characterization device is convenient to build, easy to operate, and has a wide range of applications. Combined with the in-situ characterization method of the present invention, it realizes the principle of interaction between microorganisms and solid surfaces in the bright field of an inverted microscope without labeling microorganisms. Position, real-time, and long-term observation can be effectively used for the study of the adhesion kinetics and proliferation behavior of microorganisms on solid surfaces.

Description

technical field [0001] The invention belongs to the interdisciplinary technical fields of microscopy, microbiology and materials science, and specifically relates to an in-situ characterization device and a characterization method for the interaction between microorganisms and solid surfaces. Background technique [0002] Microorganisms are everywhere and are closely related to human beings. The existence of beneficial microbial systems not only determines the health of the human body (such as intestinal flora), but also plays an important role that other substances cannot replace in wastewater treatment, metal recovery, and fine chemical preparation. However, while bringing convenience to human production and life, problems such as high energy consumption caused by the growth of microorganisms on the surfaces of food packaging, shipping, biological materials, and implanted devices have also brought heavy economic burdens to humans, and even Direct threats to human life saf...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/84G01N21/01
Inventor 杨小牛高沁薇陈兆彬李金歌
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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