CN112871229B - Chip for water dielectrophoresis bacteria sorting - Google Patents
Chip for water dielectrophoresis bacteria sorting Download PDFInfo
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- CN112871229B CN112871229B CN202110088291.4A CN202110088291A CN112871229B CN 112871229 B CN112871229 B CN 112871229B CN 202110088291 A CN202110088291 A CN 202110088291A CN 112871229 B CN112871229 B CN 112871229B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
- C12M47/02—Separating microorganisms from the culture medium; Concentration of biomass
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/50273—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by the means or forces applied to move the fluids
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502761—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/08—Geometry, shape and general structure
- B01L2300/0887—Laminated structure
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
- B01L2400/0424—Dielectrophoretic forces
Abstract
The invention relates to a chip for dielectrophoresis bacteria sorting of a water body, which comprises a dielectrophoresis bacteria sorting chip module, wherein the dielectrophoresis bacteria sorting chip module comprises an intracavity structure and an extracavity structure; the extraluminal structure comprises a microchannel; the cavity structure is of a multilayer structure and comprises an electrode layer, an insulating layer and a cavity channel structure; the electrode layer is a trapezoidal microstructure; the outlet of the cavity structure is divided into two paths, one path is directly connected to the sample waste liquid outlet through the micro-channel, and the other path is directly connected to the liquid inlet of the impedance bacteria testing module through the micro-channel. The invention introduces dielectrophoresis technology into the micro-fluidic chip, can realize the rapid collection of bacteria in the channel to the detection site within the second level, integrates the impedance electrode as the detection electrode of the bacteria quantity in the micro-fluidic chip at the same time, and has the advantages of rapid response, high precision and the like.
Description
Technical Field
The invention relates to the technical field of environmental detection, in particular to a chip for water dielectrophoresis bacteria sorting.
Background
Water is a source of life, and the safety of national drinking water is guaranteed. In the traditional chemical laboratory method in the national standard GB5750-85, a plate culture method is adopted, a water sample is subjected to microbial culture for 24-48 hours, and then the colony count on a plate is observed by naked eyes. It is therefore necessary to establish a more comprehensive and faster detection method.
Some foreign large-scale instrument companies adopt large-scale instruments such as flow cytometers, fluorescence biochemical luminometers and the like, the instruments are extremely expensive and large in size, the instruments cannot enter thousands of households, most of used reagents are not friendly to the environment, and secondary pollution is caused for a large number of civilians. Therefore, there is a great need for a portable rapid-detection instrument.
The micro-fluidic chip is a core technology platform, the main technical background of the micro-fluidic chip is the micro-fluidic technology, one or more micro-liquids are processed and operated by constructing channels with the diameter of several micrometers to hundreds of micrometers, and the micro-fluidic chip is formed by mutually communicating a series of complex micro basic units. The microfluidics technology is introduced into the project as a core technology platform due to the advantages of small sample demand, rapid analysis, easy portability, matching with the size of bacteria and the like.
Disclosure of Invention
The invention provides a chip for dielectrophoresis bacteria separation of a water body, which can solve the technical problem.
In order to achieve the purpose, the invention adopts the following technical scheme:
the dielectrophoresis bacteria sorting chip module comprises a cavity structure and an outer cavity structure;
the extraluminal structure comprises a microchannel;
the cavity structure is of a multilayer structure and comprises an electrode layer, an insulating layer and a cavity channel structure, wherein the insulating layer is arranged on the electrode layer, and the cavity channel structure is arranged on the insulating layer;
the electrode layers are two opposite microelectrodes, and trapezoidal microstructures are arranged on the microelectrodes; two microelectrodes of the electrode layer are respectively connected to two terminals of the excitation circuit module;
the outlet of the cavity structure is divided into two paths, one path is directly connected to a sample waste liquid outlet through a micro-channel, and the other path is that the liquid outlet of the dielectrophoresis module is directly connected to the liquid inlet of the impedance bacteria test module through a micro-channel.
Furthermore, the impedance bacteria test module also comprises a micro-channel layer and an electrode layer, wherein the electrode layer comprises an excitation electrode and a receiving electrode, and the excitation electrode and the receiving electrode are respectively connected to the excitation circuit module and the detection circuit module.
Furthermore, the dielectrophoresis bacteria sorting chip module is provided with a sample inlet channel and a buffer solution inlet channel, the sample inlet channel is directly connected to the front end of the dielectrophoresis bacteria sorting electrode cavity, and the buffer solution inlet channel is connected to the front end of the electrode cavity and converged to enter the front end of the dielectrophoresis bacteria sorting electrode cavity.
Furthermore, the electrode layer is composed of a pair of opposite microstructures respectively provided with plane protrusions, and the arrangement mode of the protruding microstructures is an opposite type and a staggered type.
Furthermore, the shape of the protrusion is one of a rectangle, a semicircle, a triangle and a trapezoid.
Furthermore, electromagnetic valves are arranged on the micro-channels directly connected with the dielectrophoresis bacteria sorting chip module and the impedance bacteria testing chip module.
Further, the electromagnetic valve is connected with an upper computer.
Furthermore, the electrode layer is made of Au of 50-100nm and Cr or Ti of 5-10 nm.
Further, the thickness of the insulating layer should be more than 50 μm and less than 200 μm.
According to the technical scheme, the chip for water dielectrophoresis bacteria sorting is used for developing rapid detection of bacteria number, and is a set of chip for detecting the bacteria number in water on site. The invention realizes the whole process from quick collection to quick detection of bacteria in water body in a channel with the width and the height of dozens of microns by optimizing the structure and the structural parameters of the micro-fluidic chip, and has the characteristics of high integration, high precision and the like. The dielectrophoresis technology refers to that non-charged particles (dielectric medium) directionally move under the action of electric field, when the electrically neutral cell is placed in the non-uniform electric field, the surface charge of the cell can be respectively moved towards two ends of the cell under the action of electric field polarization to form an electric dipole, and because the electric field strength of positive and negative charges of said electric dipole is different, the cell can be acted by electric field force along a certain direction, said electric field force is dielectrophoresis force. Dielectrophoresis is particularly suitable for being combined with microfluid, and can form larger electric field gradient in a micro area through the construction of a microelectrode, the required applied voltage is lower, and the generation of bubbles and joule heat is avoided.
The invention introduces dielectrophoresis technology into the microfluidic chip, can realize the rapid collection of bacteria in the channel to the detection site within the second level, the thickness of the micro electrode is only hundred nanometers level, which is equivalent to the size order of the bacteria, and the bacteria can be rapidly and completely collected at the detection site as far as possible by optimizing the control parameters. And an impedance electrode is simultaneously integrated in the microfluidic chip to serve as a detection electrode for the number of bacteria, so that the detection advantages of quick response, high precision and the like are achieved.
Drawings
FIG. 1 is a schematic structural view of the present invention;
FIG. 2 is a schematic view of the internal structure of the present invention;
FIG. 3 is a schematic structural diagram of an impedance bacteria test module of the present invention;
fig. 4 is a schematic structural view of an electrode layer of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention.
When the chip for dielectrophoresis bacteria sorting of the water body is used for detecting water quality, the chip can be combined with a control module, an electronic valve and other devices to realize automatic control;
as shown in fig. 1 and 2, the dielectrophoresis bacteria sorting chip module is included, and comprises an intracavity structure and an extracavity structure; the extraluminal structure comprises a microchannel;
the cavity structure is of a multilayer structure and comprises an electrode layer, an insulating layer and a cavity channel structure, wherein the insulating layer is arranged on the electrode layer, and the cavity channel structure is arranged on the insulating layer;
the electrode layers are two opposite microelectrodes, and trapezoidal microstructures are arranged on the microelectrodes; two microelectrodes of the electrode layer are respectively connected to two terminals of the excitation circuit module;
the outlet of the cavity structure is divided into two paths, one path is directly connected to the sample waste liquid outlet through the micro-channel, and the other path is directly connected to the liquid inlet of the impedance bacteria testing module through the micro-channel.
The sample inlet channel is directly connected to the front end of the dielectrophoresis bacteria sorting electrode cavity, and the buffer solution inlet channel is connected to the front end of the electrode cavity and converged to enter the front end of the dielectrophoresis bacteria sorting electrode cavity.
The micro-channel is mainly arranged outside the cavity, the multi-layer structure is arranged in the cavity and comprises an electrode layer, the electrode layer is mainly characterized by convex rectangular, semicircular, triangular and trapezoidal microstructures, and the arrangement mode is opposite and staggered. As shown in fig. 4, the electrode layer of the present example provides 8 cases.
The outlet of the cavity is divided into two paths, one path is directly connected to the sample waste liquid outlet, and the other path is the liquid outlet of the dielectrophoresis module and is also directly connected to the liquid inlet of the impedance module.
The principle is as follows: the bacteria are influenced by two forces of fluid force and dielectrophoresis force in the micro-channel, the fluid force is mainly controlled by flow rate, the dielectrophoresis force is determined by the convex structure and an applied alternating current signal, and the bacteria move towards the path of the inlet of the impedance module under the influence of the two forces after entering the sorting cavity.
As shown in fig. 3, the electrophoresis chip and the impedance bacteria test chip module also include electrode layers. The electrode structure is shown in fig. 4. The electrode layer comprises an excitation electrode and a receiving electrode, and the excitation electrode and the receiving electrode are respectively connected to the excitation circuit module and the detection circuit module.
The micro-channel between the dielectrophoresis bacteria sorting chip module and the impedance bacteria testing chip module can be provided with an electromagnetic valve, and the electromagnetic valve is connected with an upper computer to realize automatic control.
The height of the micro-channel of the embodiment of the invention is more than 100 mu m, the micro-fluidic channel layer is also made of polymer plastic and is an elastic body, and the elastic body forms the micro-channel which is communicated with each other through a certain structural shape and is used for the liquid to circulate in the micro-channel.
The micro-fluidic channel substrate insulating layer and the micro-channel are made of the same material, so that bonding connection is facilitated. The thickness of the insulating layer should be greater than 50 μm and less than 200 μm.
The electrode layer is buried under the insulating layer and closely attached to the substrate fixing layer. The electrode layer is made of Au of 50-100nm and Cr or Ti of 5-10 nm. The electrodes are only present at specific locations.
In summary, the microfluidic chip is a chip for dielectrophoresis bacteria sorting in water, and the main technical background of the microfluidic chip is the microfluidic technology, wherein one or more trace liquids are processed and operated by constructing channels of several micrometers to hundreds of micrometers, and the microfluidic chip is formed by communicating a series of complex micro basic units. The microfluidics technology is introduced into the project as a core technology platform due to the advantages of small sample demand, rapid analysis, easy portability, matching with the size of bacteria and the like. The invention realizes the whole process from quick collection to quick detection of bacteria in water body in a channel with the width and the height of dozens of microns by optimizing the structure and the structural parameters of the micro-fluidic chip, and has the characteristics of high integration, high precision and the like. The dielectrophoresis technology refers to that non-charged particles (dielectric medium) directionally move under the action of electric field, when the electrically neutral cell is placed in the non-uniform electric field, the surface charge of the cell can be respectively moved towards two ends of the cell under the action of electric field polarization to form an electric dipole, and because the electric field strength of positive and negative charges of said electric dipole is different, the cell can be acted by electric field force along a certain direction, said electric field force is dielectrophoresis force. Dielectrophoresis is particularly suitable for being combined with microfluid, and can form larger electric field gradient in a micro-area through the construction of a microelectrode, the required applied voltage is lower, and the generation of bubbles and joule heat is avoided.
The embodiment of the invention introduces a dielectrophoresis technology into the microfluidic chip, can realize the rapid collection of bacteria in a channel to a detection site within a second level, has the thickness of a microelectrode of hundreds of nanometers level which is equivalent to the size level of the bacteria, and ensures that the bacteria are rapidly and completely collected at the detection site as far as possible by optimizing control parameters. And an impedance electrode is simultaneously integrated in the microfluidic chip to serve as a detection electrode for the number of bacteria, so that the detection advantages of quick response, high precision and the like are achieved.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (5)
1. A chip for dielectrophoresis bacteria separation of a water body is characterized in that:
the dielectrophoresis bacteria sorting chip module comprises an intracavity structure and an extracavity structure;
the extraluminal structure comprises a microchannel;
the cavity structure is of a multilayer structure and comprises an electrode layer, an insulating layer and a cavity channel structure, wherein the insulating layer is arranged on the electrode layer, and the cavity channel structure is arranged on the insulating layer;
the electrode layers are two opposite microelectrodes, and trapezoidal microstructures are arranged on the microelectrodes; two microelectrodes of the electrode layer are respectively connected to two terminals of the excitation circuit module;
the outlet of the cavity structure is divided into two paths, one path is directly connected to a sample waste liquid outlet through a micro-channel, and the other path is that a dielectrophoresis module liquid outlet is directly connected to an impedance bacteria test module liquid inlet through a micro-channel;
the impedance bacteria test module also comprises a micro-channel layer and an electrode layer, wherein the electrode layer comprises an excitation electrode and a receiving electrode, and the excitation electrode and the receiving electrode are respectively connected to the excitation circuit module and the detection circuit module;
the dielectrophoresis bacteria sorting chip module is provided with a sample inlet channel and a buffer solution inlet channel, the sample inlet channel is directly connected to the front end of the dielectrophoresis bacteria sorting electrode cavity, and the buffer solution inlet channel is connected to the front end of the electrode cavity and enters the front end of the dielectrophoresis bacteria sorting electrode cavity after being converged;
the electrode layer is composed of a pair of opposite microstructures respectively provided with plane protrusions, and the arrangement mode of the protruding microstructures is an opposite type and a staggered type;
electromagnetic valves are arranged on the micro-channels directly connected with the dielectrophoresis bacteria sorting chip module and the impedance bacteria testing chip module.
2. The chip for dielectrophoresis of bacteria in a water body according to claim 1, wherein: the shape of the protruding microstructure is one of rectangle, semicircle, triangle and trapezoid.
3. The chip for dielectrophoresis of bacteria in a water body according to claim 1, wherein: the electromagnetic valve is connected with an upper computer.
4. The dielectrophoresis bacteria sorting chip for a water body according to claim 1, wherein: the electrode layer is made of 50-100nm Au and 5-10nm Cr or Ti.
5. The chip for dielectrophoresis of bacteria in a water body according to claim 1, wherein: the thickness of the insulating layer should be greater than 50 μm and less than 200 μm.
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