CN113289701A - Pump-free micro-fluidic chip for electrochemical detection and preparation method thereof - Google Patents
Pump-free micro-fluidic chip for electrochemical detection and preparation method thereof Download PDFInfo
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- CN113289701A CN113289701A CN202110554831.3A CN202110554831A CN113289701A CN 113289701 A CN113289701 A CN 113289701A CN 202110554831 A CN202110554831 A CN 202110554831A CN 113289701 A CN113289701 A CN 113289701A
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- 238000000835 electrochemical detection Methods 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title abstract description 9
- 239000012530 fluid Substances 0.000 claims abstract description 47
- 238000006243 chemical reaction Methods 0.000 claims abstract description 39
- 238000001514 detection method Methods 0.000 claims abstract description 27
- AMGQUBHHOARCQH-UHFFFAOYSA-N indium;oxotin Chemical compound [In].[Sn]=O AMGQUBHHOARCQH-UHFFFAOYSA-N 0.000 claims abstract description 23
- 238000002347 injection Methods 0.000 claims description 12
- 239000007924 injection Substances 0.000 claims description 12
- 238000002156 mixing Methods 0.000 claims description 10
- 238000004140 cleaning Methods 0.000 claims description 9
- 229920000742 Cotton Polymers 0.000 claims description 8
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 6
- 239000008367 deionised water Substances 0.000 claims description 6
- 229910021641 deionized water Inorganic materials 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 230000003647 oxidation Effects 0.000 claims description 6
- 238000007254 oxidation reaction Methods 0.000 claims description 6
- 238000004506 ultrasonic cleaning Methods 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 238000007789 sealing Methods 0.000 claims description 5
- 239000004205 dimethyl polysiloxane Substances 0.000 claims description 4
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 claims description 4
- -1 polydimethylsiloxane Polymers 0.000 claims description 4
- 238000005086 pumping Methods 0.000 claims description 4
- 238000001755 magnetron sputter deposition Methods 0.000 claims description 3
- 238000004080 punching Methods 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 3
- 239000005361 soda-lime glass Substances 0.000 claims description 3
- 239000011248 coating agent Substances 0.000 claims 1
- 238000000576 coating method Methods 0.000 claims 1
- 230000010354 integration Effects 0.000 abstract description 7
- 238000004458 analytical method Methods 0.000 abstract description 4
- 238000011527 multiparameter analysis Methods 0.000 abstract description 2
- 238000005516 engineering process Methods 0.000 description 7
- 238000003825 pressing Methods 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 238000007747 plating Methods 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000012742 biochemical analysis Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000004949 mass spectrometry Methods 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
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- 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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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
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Abstract
The invention belongs to the technical field of biochemical detection, in particular to a pump-free micro-fluidic chip for electrochemical detection and a preparation method thereof, which solve the problems that external driving equipment for driving the fluid of the micro-fluidic chip in the prior art is complex and heavy, cannot be carried about, and is not easy to realize miniaturization and integration; the chip body is provided with a sample feeding area, a pre-reaction area, a mixed reaction area, a detection area, a capillary area and a finger-pressure pump area which are sequentially connected through a connecting channel; the Indium Tin Oxide (ITO) electrode is a three-electrode system and comprises a working electrode, a counter electrode and a reference electrode. The pump-free micro-fluidic chip provided by the invention has high integration level, and is very suitable for multi-parameter analysis in various integrated, portable and miniaturized analysis and detection.
Description
Technical Field
The invention relates to the technical field of biochemical detection, in particular to a pump-free micro-fluidic chip for electrochemical detection and a preparation method thereof.
Background
In recent years, microfluidic technology has been developed rapidly, and has been widely studied and applied in various fields such as biology, chemistry, and medicine. Microfluidic chips enable the integration of traditional conventional laboratory operations (such as sample introduction, mixing, reaction, extraction, separation, detection, etc.) onto chips of only a few square centimeters in size, reducing analysis time from hours to tens of seconds or even less. At the present stage, the contribution of the microfluidic technology in the aspect of realizing 'on-site instant detection' has important significance on the perfection and health of global public health service.
The detection for the micro-fluidic chip mainly comprises optical detection, electrochemical detection, mass spectrometry detection and the like at present, and the electrochemical detection technology has higher sensitivity, so that the used electrode is easy to realize miniaturization and integration, the detection sensitivity is not reduced, and the electrochemical detection technology has the advantages of no optical path, low price and the like, so that the electrochemical detection technology has incomparable advantages compared with other detection technologies in the aspect of constructing a miniaturized, portable and integrated micro-fluidic system.
The rapid development of microfluidic chips also puts higher demands on the fluid driving manner. At present, injection pumps, pneumatic pumps, electric drives, micro valves or micro pumps and the like are mainly used for driving the fluid of the microfluidic chip, however, the complex and heavy external driving equipment cannot be carried about, and is not easy to realize miniaturization and integration, so that the pump-free microfluidic chip which is designed to realize miniaturization, is convenient to integrate, has controllable flow direction in a chip channel and does not influence the original characteristics of organisms and the working stability of an analysis system has great significance. Based on the statement, the invention provides a pump-free micro-fluidic chip for electrochemical detection and a preparation method thereof.
Disclosure of Invention
The invention aims to solve the problems that external driving equipment for driving a microfluidic chip fluid in the prior art is complex and heavy, cannot be carried about, and is not easy to realize miniaturization and integration, and provides a pump-free microfluidic chip for electrochemical detection and a preparation method thereof.
A pump-free micro-fluidic chip for electrochemical detection comprises a chip body and an Indium Tin Oxide (ITO) electrode fixedly attached to the bottom of the chip body; the chip body is provided with a sample feeding area, a pre-reaction area, a mixed reaction area, a detection area, a capillary area and a finger-pressure pump area which are sequentially connected through a connecting channel; the sample feeding area and the pre-reaction area are arranged in the left-right direction and are close to the front end of the chip body, the detection area is positioned in the middle of the chip body, and the capillary area and the finger-pressure pump area are close to the rear end of the chip body; the Indium Tin Oxide (ITO) electrode is a three-electrode system and comprises a working electrode, a counter electrode and a reference electrode.
Preferably, the sample injection region comprises at least two sample injection holes which vertically penetrate through the chip body, and the diameter of each sample injection hole is 2-6 mm.
Preferably, the pre-reaction zone comprises a pre-reaction tank, at least two sample inlet holes of the sample inlet zone are respectively merged at the pre-reaction tank through at least two first connecting channels, and the diameter of the pre-reaction tank is 1-3 mm.
Preferably, the mixing reaction zone comprises a tortuous fluid channel I, the inlet end of the fluid channel I is connected with the outlet end of the pre-reaction tank through a second connecting channel, and the outlet end of the fluid channel I is connected with the inlet end of the detection zone through a third connecting channel.
Preferably, the detection zone comprises a circuitous and curved fluid channel II, and the fluid channel II in the detection zone is longer and more compact than the fluid channel I in the mixing reaction zone, the inlet end of the fluid channel II is connected with the outlet end of the mixing reaction zone through a third connecting channel, and the outlet end of the fluid channel II is connected with the inlet end of the capillary zone through a fourth connecting channel.
Preferably, the capillary zone includes a plurality of capillary shunt passages, an axial direction of each capillary passage extends in a front-rear direction, the plurality of capillary shunt passages integrally present a fan shape, and the plurality of capillary shunt passages are arranged side by side in a horizontal plane.
Preferably, the finger-pressure pump area is in a fan shape, no fluid channel is arranged in the finger-pressure pump area, and a channel is arranged outside the fan shape to define the finger-pressure pump area for placing absorbent cotton before chip sealing.
Preferably, the height of the chip body is 4-8mm, the length of the chip body in the front-back direction is 45-55mm, the diameter of the sample inlet is 2-6mm, and the widths of the fluid channel I, the fluid channel II, the plurality of capillary channels, the first connecting channel, the second connecting channel, the third connecting channel and the fourth connecting channel are 200-500 um.
Preferably, the Indium Tin Oxide (ITO) electrode is a three-electrode, and comprises a working electrode, a counter electrode, and a reference electrode, wherein the working electrode is located in the middle, the counter electrode is located on the right, and the reference electrode is located on the left, and the Indium Tin Oxide (ITO) electrode is fabricated by plating a transparent Indium Tin Oxide (ITO) film on soda-lime glass by a magnetron sputtering method.
The invention also provides a preparation method of the pump-free micro-fluidic chip for electrochemical detection, which comprises the following steps:
(1) selecting polydimethylsiloxane as a raw material, firstly processing a chip blank of the pump-free microfluidic chip, forming a corresponding reaction groove, a fluid channel, a plurality of capillary channels, a first connecting channel, a second connecting channel, a third connecting channel and a fourth connecting channel at the bottom of the chip blank, then punching holes at corresponding positions on the chip blank to form sample inlet holes of the pump-free microfluidic chip, and taking the punched chip blank as a chip body;
(2) taking a clean Indium Tin Oxide (ITO) electrode for standby;
(3) the chip body is cleaned and dried, and the cleaning operation is as follows: firstly, putting a chip body into a container containing ethanol, placing the container in an ultrasonic cleaning machine, cleaning for 5-8 minutes, taking out the chip body, washing the chip body with deionized water for 5-8 times, then putting the chip body into the container containing isopropanol, placing the container in the ultrasonic cleaning machine, cleaning for 5-8 minutes, taking out the chip body, and washing the chip body with deionized water for 5-8 times;
(4) placing required absorbent cotton in a finger-pressure pump area of the chip body;
(5) and (3) carrying out plasma oxidation on the chip body for 90s, and immediately carrying out firm sealing on the chip body and the Indium Tin Oxide (ITO) electrode in the step (2) after the oxidation is finished, thus obtaining a pump-free micro-fluidic chip finished product.
The pump-free micro-fluidic chip for electrochemical detection provided by the invention has the following beneficial effects:
1. the pump-free microfluidic chip for electrochemical detection, provided by the invention, has the advantages that the height of a chip body is 4-8mm, the length of the chip body in the front-back direction is 45-55mm, the diameter of a sample inlet is 2-6mm, and the widths of a fluid channel I, a fluid channel II, a plurality of capillary channels, a first connecting channel, a second connecting channel, a third connecting channel and a fourth connecting channel are 200-500 um; small volume, convenient carrying and being beneficial to realizing instant detection.
2. The pump-free micro-fluidic chip for electrochemical detection provided by the invention really realizes a pump-free technology without using a complex external pumping system. The required absorbent cotton is placed in the finger-pressure pump area of the chip body in advance, the driving pressure of fingers is utilized to provide initial power for fluid in the pump-free micro-fluidic chip channel, and when the fluid flows through the capillary area, continuous power is provided for the fluid in the pump-free micro-fluidic chip channel by utilizing the capillary force of the capillary channel and the absorbent cotton. The pump-free micro-fluidic chip provided by the invention has high integration level, and the steps of sample adding, mixing, reacting, detecting and the like of a sample are integrated on a chip with a square centimeter, so that the pump-free micro-fluidic chip is very suitable for multi-parameter analysis in various integrated, portable and miniaturized analysis and detection.
3. According to the preparation method of the pump-free microfluidic chip for electrochemical detection, disclosed by the invention, the polydimethylsiloxane is used as the blank to process the pump-free microfluidic chip, and the pump-free microfluidic chip has the excellent characteristics of good biocompatibility, light transmittance, low price, simple use method and the like, and is very suitable for biochemical analysis and detection.
Drawings
Fig. 1 is a schematic structural diagram of a chip body of a pump-free microfluidic chip that can be used for electrochemical detection according to the present invention.
Fig. 2 is a schematic structural diagram of an Indium Tin Oxide (ITO) electrode of a pump-free microfluidic chip for electrochemical detection according to the present invention.
Detailed Description
The embodiments of the present invention will be described more fully hereinafter with reference to the accompanying drawings.
Examples
The following can be understood by combining the attached drawings 1 and 2: the invention discloses a pump-free micro-fluidic chip, which comprises a chip body and an Indium Tin Oxide (ITO) electrode fixedly attached to the bottom of the chip body, wherein a sample feeding area, a pre-reaction area, a mixed reaction area, a detection area, a capillary area and a finger-pressure pump area which are sequentially connected through a connecting channel are arranged on the chip body;
the sample injection region comprises at least two sample injection holes, the sample injection holes vertically penetrate through the chip body of the chip, and the diameters of the sample injection holes are 2-6 mm;
the pre-reaction zone comprises a pre-reaction tank, at least two sample inlet holes of the sample inlet zone are respectively converged at the pre-reaction tank through at least two first connecting channels, and the diameter of the pre-reaction tank is 1-3 mm;
the mixed reaction zone comprises a roundabout and bent fluid channel I, the inlet end of the fluid channel I is connected with the outlet end of the pre-reaction tank through a second connecting channel, and the outlet end of the fluid channel I is connected with the inlet end of the detection zone through a third connecting channel;
the detection area comprises a circuitous and bent fluid channel II, and compared with the fluid channel I of the mixing reaction area, the fluid channel II of the detection area is longer and more compact, the inlet end of the fluid channel II is connected with the outlet end of the mixing reaction area through a third connecting channel, and the outlet end of the fluid channel II is connected with the inlet end of the capillary area through a fourth connecting channel;
the capillary area comprises a plurality of capillary shunt channels, the axial direction of each capillary channel extends along the front-back direction, the plurality of capillary shunt channels are integrally in a fan shape, and the plurality of capillary shunt channels are arranged side by side in the horizontal plane;
the finger pressing pump area is in a fan shape, no fluid channel is arranged in the finger pressing pump area, and a channel is arranged outside the fan shape to define the finger pressing pump area and place absorbent cotton before the chip is sealed;
the pre-reaction tank, the fluid channel I, the fluid channel II, the plurality of capillary channels, the first connecting channel, the second connecting channel, the third connecting channel and the fourth connecting channel are all arranged at the bottom of the chip body and are tightly sealed by Indium Tin Oxide (ITO) electrodes;
the height of the chip body is 4-8mm, the length of the chip body in the front-back direction is 45-55mm, the diameter of the sample inlet is 2-6mm, and the widths of the fluid channel I, the fluid channel II, the plurality of capillary channels, the first connecting channel, the second connecting channel, the third connecting channel and the fourth connecting channel are 200-500 um;
the Indium Tin Oxide (ITO) electrode is a three-electrode system and comprises a working electrode, a counter electrode and a reference electrode, wherein the working electrode is positioned in the middle, the counter electrode is positioned on the right side, the reference electrode is positioned on the left side, and the Indium Tin Oxide (ITO) electrode is manufactured by plating a layer of transparent Indium Tin Oxide (ITO) film on soda-lime glass by a magnetron sputtering method.
The invention also provides a preparation method of the pump-free micro-fluidic chip for electrochemical detection, which comprises the following steps:
(1) selecting polydimethylsiloxane as a raw material, firstly processing a chip blank of the pump-free microfluidic chip, forming a corresponding reaction groove, a fluid channel, a plurality of capillary channels, a first connecting channel, a second connecting channel, a third connecting channel and a fourth connecting channel at the bottom of the chip blank, then punching holes at corresponding positions on the chip blank to form sample inlet holes of the pump-free microfluidic chip, and taking the punched chip blank as a chip body;
(2) taking a clean Indium Tin Oxide (ITO) electrode for standby;
(3) the chip body is cleaned and dried, and the cleaning operation is as follows: firstly, putting a chip body into a container containing ethanol, placing the container in an ultrasonic cleaning machine, cleaning for 5-8 minutes, taking out the chip body, washing the chip body with deionized water for 5-8 times, then putting the chip body into the container containing isopropanol, placing the container in the ultrasonic cleaning machine, cleaning for 5-8 minutes, taking out the chip body, and washing the chip body with deionized water for 5-8 times;
(4) placing required absorbent cotton in a finger-pressure pump area of the chip body;
(5) and (3) carrying out plasma oxidation on the chip body for 90s, and immediately carrying out firm sealing on the chip body and the Indium Tin Oxide (ITO) electrode in the step (2) after the oxidation is finished, thus obtaining a pump-free micro-fluidic chip finished product.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (10)
1. A pump-free micro-fluidic chip for electrochemical detection is characterized by comprising a chip body and an indium tin oxide electrode fixedly attached to the bottom of the chip body; the chip body is provided with a sample feeding area, a pre-reaction area, a mixed reaction area, a detection area, a capillary area and a finger-pressure pump area which are sequentially connected through a connecting channel; the sample feeding area and the pre-reaction area are arranged in the left-right direction and are close to the front end of the chip body, the detection area is positioned in the middle of the chip body, and the capillary area and the finger-pressure pump area are close to the rear end of the chip body; the indium tin oxide electrode is a three-electrode system and comprises a working electrode, a counter electrode and a reference electrode.
2. The pump-free microfluidic chip for electrochemical detection according to claim 1, wherein the sample injection region comprises at least two sample injection holes, the sample injection holes penetrate through the chip body from top to bottom, and the diameter of each sample injection hole is 2-6 mm.
3. The pump-free microfluidic chip for electrochemical detection according to claim 1, wherein the pre-reaction region comprises a pre-reaction groove, at least two sample inlets of the sample inlet region are respectively merged at the pre-reaction groove through at least two first connecting channels, and the diameter of the pre-reaction groove is 1-3 mm.
4. The pump-free microfluidic chip for electrochemical detection according to claim 1, wherein the mixing reaction area comprises a tortuous fluid channel I, an inlet end of the fluid channel I is connected to an outlet end of the pre-reaction tank through a second connecting channel, and an outlet end of the fluid channel I is connected to an inlet end of the detection area through a third connecting channel.
5. The pump-free microfluidic chip for electrochemical detection according to claim 1, wherein the detection region comprises a tortuous fluid channel II, and the fluid channel II is longer and more compact than the fluid channel I of the mixing reaction region, the inlet end of the fluid channel II is connected to the outlet end of the mixing reaction region through a third connecting channel, and the outlet end of the fluid channel II is connected to the inlet end of the capillary region through a fourth connecting channel.
6. The pump-free microfluidic chip according to claim 1, wherein the capillary region comprises a plurality of capillary shunt channels, each of the capillary channels extends in a front-back direction in an axial direction, the plurality of capillary shunt channels are integrally fan-shaped, and the plurality of capillary shunt channels are arranged side by side in a horizontal plane.
7. The pump-free microfluidic chip for electrochemical detection according to claim 1, wherein the acupressure pumping region is a sector shape, no fluid channel is provided in the acupressure pumping region, and a channel is provided outside the sector shape to define the acupressure pumping region for placing absorbent cotton before sealing the chip.
8. The pump-free microfluidic chip for electrochemical detection as claimed in claim 1, wherein the height of the chip body is 4-8mm, the length of the chip body in the front-back direction is 45-55mm, the diameter of the sample inlet is 2-6mm, and the widths of the fluid channel I, the fluid channel II, the plurality of capillary channels, the first connecting channel, the second connecting channel, the third connecting channel and the fourth connecting channel are 200-500 um.
9. The pump-free microfluidic chip for electrochemical detection according to claim 1, wherein the indium tin oxide electrode is a three-electrode structure comprising a working electrode, a counter electrode and a reference electrode, the working electrode is located in the middle, the counter electrode is located on the right side, and the reference electrode is located on the left side, and the indium tin oxide electrode is formed by coating a transparent indium tin oxide film on soda-lime glass by a magnetron sputtering method.
10. A method for preparing a pump-free microfluidic chip for electrochemical detection according to any one of claims 1 to 9, comprising the following steps:
(1) selecting polydimethylsiloxane as a raw material, firstly processing a chip blank of the pump-free microfluidic chip, forming a corresponding reaction groove, a fluid channel, a plurality of capillary channels, a first connecting channel, a second connecting channel, a third connecting channel and a fourth connecting channel at the bottom of the chip blank, then punching holes at corresponding positions on the chip blank to form sample inlet holes of the pump-free microfluidic chip, and taking the punched chip blank as a chip body;
(2) taking a clean indium tin oxide electrode for standby;
(3) the chip body is cleaned and dried, and the cleaning operation is as follows: firstly, putting a chip body into a container containing ethanol, placing the container in an ultrasonic cleaning machine, cleaning for 5-8 minutes, taking out the chip body, washing the chip body with deionized water for 5-8 times, then putting the chip body into the container containing isopropanol, placing the container in the ultrasonic cleaning machine, cleaning for 5-8 minutes, taking out the chip body, and washing the chip body with deionized water for 5-8 times;
(4) placing required absorbent cotton in a finger-pressure pump area of the chip body;
(5) and (3) carrying out plasma oxidation on the chip body for 90s, and immediately carrying out firm sealing on the chip body and the indium tin oxide electrode in the step (2) after the oxidation is finished, thus obtaining a pump-free micro-fluidic chip finished product.
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