CN114471752A - Chip and preparation method thereof - Google Patents
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- CN114471752A CN114471752A CN202011167194.6A CN202011167194A CN114471752A CN 114471752 A CN114471752 A CN 114471752A CN 202011167194 A CN202011167194 A CN 202011167194A CN 114471752 A CN114471752 A CN 114471752A
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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
- B01L3/502707—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 manufacture of the container or its components
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B81—MICROSTRUCTURAL TECHNOLOGY
- B81C—PROCESSES OR APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OR TREATMENT OF MICROSTRUCTURAL DEVICES OR SYSTEMS
- B81C1/00—Manufacture or treatment of devices or systems in or on a substrate
- B81C1/00015—Manufacture or treatment of devices or systems in or on a substrate for manufacturing microsystems
- B81C1/00023—Manufacture or treatment of devices or systems in or on a substrate for manufacturing microsystems without movable or flexible elements
- B81C1/00119—Arrangement of basic structures like cavities or channels, e.g. suitable for microfluidic systems
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B81—MICROSTRUCTURAL TECHNOLOGY
- B81C—PROCESSES OR APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OR TREATMENT OF MICROSTRUCTURAL DEVICES OR SYSTEMS
- B81C1/00—Manufacture or treatment of devices or systems in or on a substrate
- B81C1/00436—Shaping materials, i.e. techniques for structuring the substrate or the layers on the substrate
- B81C1/00523—Etching material
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B81—MICROSTRUCTURAL TECHNOLOGY
- B81C—PROCESSES OR APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OR TREATMENT OF MICROSTRUCTURAL DEVICES OR SYSTEMS
- B81C3/00—Assembling of devices or systems from individually processed components
- B81C3/001—Bonding of two components
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
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Abstract
The application relates to the field of magnetic control micro-fluidic chips and discloses a chip and a preparation method thereof, wherein the chip comprises: the glass substrate, soft magnetic metal layer and micro-fluidic channel, wherein, micro-fluidic channel and glass substrate bonding are so that soft magnetic metal layer is located between glass substrate and micro-fluidic channel. The chip disclosed by the application improves the capture rate of magnetic beads in the nucleic acid extraction process and improves the experimental recovery rate by arranging the soft magnetic metal layer between the glass substrate and the microfluidic channel. In addition, the integrated design reduces the dependence on an external magnetic field, so that the adaptability of the chip and the device is higher.
Description
Technical Field
The application relates to the field of magnetic control micro-fluidic chips, in particular to a chip and a preparation method thereof.
Background
The magnetic material is one of the shapes of the microbeads, and is widely applied to the fields of biological separation, targeted drug loading, nuclear magnetic resonance imaging and the like due to the characteristics of large surface area, good magnetic guidance, strong operability and the like. The magnetic control micro-fluidic chip can be used for mixing and transporting liquid in the micro-fluidic chip, switching and valving of the chip, and transporting, separating and capturing magnetic objects.
The molecular diagnosis uses nucleic acid as a detection object, is mainly applied to diagnosis of clinical departments, such as tumors, infectious diseases, heredity and the like, and nucleic acid extraction is the 'first step' of the molecular diagnosis. Magnetic beads are a biological functional material based on nanotechnology, and are prepared by wrapping magnetic nanoparticles into microspheres and performing surface treatment (biochemical modification) to enable the microspheres to be combined with various target detection substances. The DNA/RNA is released from the cells or tissues under the action of the lysis solution and is specifically combined with the modified magnetic beads to form a magnetic bead-nucleic acid compound. Under the action of an external magnetic field, when the magnetic bead solution flows through the microchannel at a certain flow speed, the magnetic bead solution is acted by the magnetic field force and is adsorbed and captured. When the external magnetic field is removed, the magnetic beads are released and re-suspended in the solution, so that the separation and purification of the nucleic acid are realized.
At present, the magnetic field in the microfluidic chip is mainly generated by an external magnet or an integrated soft magnet, the conventional permanent magnet or electromagnet or soft magnet is placed outside the microfluidic chip, and magnetic particles are controlled by the magnetic field intensity of the conventional permanent magnet or electromagnet or soft magnet. Therefore, how to increase the capture rate of magnetic beads in a nucleic acid extraction process of a microfluidic chip is a problem to be solved by those skilled in the art.
Disclosure of Invention
The invention provides a chip, which improves the capture rate of magnetic beads in the nucleic acid extraction process and the experimental recovery rate by placing a soft magnet inside the chip.
In order to achieve the above object, the present invention provides a chip comprising:
the glass substrate, the soft magnetic metal layer and the microfluidic channel are bonded with the glass substrate, so that the soft magnetic metal layer is located between the glass substrate and the microfluidic channel.
According to the chip, the soft magnetic metal layer is arranged between the glass substrate and the microfluidic channel, when the magnetic beads move in the microfluidic channel, the magnetic beads are controlled to move by applying an external magnetic field, when the external magnetic field is applied, the magnetic beads are adsorbed at the bottom of the microfluidic channel, and when the external magnetic field is removed, the magnetic beads recover the suspended state, so that the magnetic beads are captured and released, and separation and purification of substances in a nucleic acid extraction experiment are completed. Because the soft magnetic metal layer is positioned in the chip, when an external magnetic field is applied, the control range of the magnetic field can be enhanced, so that the magnetic field intensity is enhanced, and the capture rate of magnetic beads is increased. In addition, the integrated design of the chip can reduce the dependence on an external magnetic field, so that the adaptability of the chip and the device is higher.
Therefore, according to the chip provided by the invention, the soft magnetic metal layer is arranged between the glass substrate and the microfluidic channel, so that the capture rate of magnetic beads in the nucleic acid extraction process is improved, and the experimental recovery rate is improved.
Preferably, the microfluidic device further comprises an insulating layer located between the soft magnetic metal layer and the microfluidic channel.
Preferably, the glass substrate further comprises a titanium seed layer and a copper seed layer which are located between the soft magnetic metal layer and the glass substrate, are sequentially arranged along the direction from the glass substrate to the soft magnetic metal layer, and correspond to the soft magnetic metal layer one by one.
Preferably, the thickness of the titanium seed layer is 0.8-1.2um, and the thickness of the copper seed layer is 0.8-1.2 um.
Preferably, along the length direction of the glass substrate, the flow channel comprises a plurality of S-shaped sub-channels which are sequentially connected end to end.
Preferably, the height of the channel is 0.5-3 mm.
Preferably, the thickness of the soft magnetic metal layer is 50 um.
Preferably, the material of the soft magnetic metal layer is one of iron-nickel alloy, pure iron and low carbon steel, iron-silicon alloy, iron-aluminum alloy, soft magnetic iron oxide ti3 and amorphous soft magnetic alloy.
Preferably, the present invention also provides a method for preparing a chip as described in any one of the above, comprising:
forming a soft magnetic metal layer on a glass substrate;
forming a microfluidic channel;
bonding the microfluidic channel to the glass substrate such that the soft magnetic metal layer is located between the glass substrate and the microfluidic channel.
Preferably, the forming of the soft magnetic metal layer on the glass substrate includes:
coating positive photoresist on a glass substrate;
developing and exposing the positive photoresist by using a patterning process to obtain a depressed part, and filling a soft magnetic metal material in the depressed part to form a soft magnetic metal layer;
and removing the residual positive photoresist.
Preferably, after removing the remaining positive photoresist, depositing a silicon oxide layer on the soft magnetic metal layer to form an insulating layer.
Preferably, before the positive photoresist is coated on the glass substrate, the method further comprises: sputtering a titanium seed layer and a copper seed layer on the glass substrate in sequence;
and when the residual positive photoresist is removed, simultaneously removing the titanium seed layer and the copper seed layer corresponding to the positive photoresist.
Preferably, the forming the microfluidic channel includes:
coating SU-8 photoresist on a silicon wafer;
developing and exposing the SU-8 photoresist by using a composition process so that the SU-8 photoresist is in the shape of a microfluidic channel;
pouring a PDMS layer on the SU-8 photoresist;
and removing the SU-8 photoresist.
Drawings
FIG. 1 is a diagram illustrating a film structure of a chip according to an embodiment of the invention;
FIG. 2 is a schematic diagram of an operating principle of a chip according to an embodiment of the present invention;
FIG. 3 is a schematic diagram of a chip according to an embodiment of the present invention;
FIG. 4 is a schematic cross-sectional view taken along the direction A in FIG. 3;
FIG. 5 is a schematic cross-sectional view taken along the direction B in FIG. 3;
FIG. 6 is a schematic diagram illustrating a step of a method for manufacturing a chip according to an embodiment of the present invention;
FIG. 7 is a diagram illustrating an embodiment of step S100 in FIG. 6;
fig. 8 is a schematic diagram of a specific step of step S200 in fig. 6.
In the figure:
10-a glass substrate; 20-a soft magnetic metal layer; 30-a titanium seed layer; 40-copper seed layer; 50-an insulating layer; 60-microfluidic channels; 70-magnetic beads; 80-PDMS.
Detailed Description
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 only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1 and fig. 3-5, the present invention provides a chip, including: a glass substrate 10, a soft magnetic metal layer 20, and a microfluidic channel 60, wherein the microfluidic channel 60 is bonded to the glass substrate 10 such that the soft magnetic metal layer 20 is located between the glass substrate 10 and the microfluidic channel 60.
In the chip, as shown in fig. 2, the soft magnetic metal layer 20 is disposed between the glass substrate 10 and the microfluidic channel 60, and by using a method of capturing the magnetic bead 70 by using a magnetic control method, the magnetic bead 70 moves in the microfluidic channel, the magnetic bead 70 is controlled to move by applying an external magnetic field, when the external magnetic field is applied, the magnetic bead 70 is adsorbed at the bottom of the microfluidic channel, and when the external magnetic field is removed, the magnetic bead 70 returns to a suspended state, so that the capture and release of the magnetic bead 70 are realized, and the separation and purification of the substances in the nucleic acid extraction experiment are completed. Since the soft magnetic metal layer 20 is located inside the chip, when an external magnetic field is applied, the control range of the magnetic field can be increased, thereby increasing the magnetic field strength, and thus increasing the capture rate of the magnetic beads 70. In addition, the integrated design of the chip can reduce the dependence on an external magnetic field, so that the adaptability of the chip and the device is higher.
Therefore, according to the chip provided by the invention, the soft magnetic metal layer 20 is arranged between the glass substrate 10 and the microfluidic channel 60, so that the capture rate of the magnetic beads 70 in the nucleic acid extraction process is improved, and the experimental recovery rate is improved.
Specifically, with continued reference to fig. 1, the chip may further include an insulating layer 50 located between the soft magnetic metal layer 20 and the microfluidic channel 60, where the insulating layer 50 may be used to protect the soft magnetic metal layer 20 to ensure the magnetic properties of the soft magnetic metal layer 20. The material of the insulating layer 50 may be silicon oxide, or may be another material.
Further, the chip may include a titanium seed layer 30 and a copper seed layer 40, which are located between the soft magnetic metal layer 20 and the glass substrate 10 and sequentially arranged in a direction from the glass substrate 10 to the soft magnetic metal layer 20, corresponding to the soft magnetic metal layer 20 one by one, and by using the titanium seed layer 30 and the copper seed layer 40, the soft magnetic metal layer 20 can be better attached to the glass substrate 10, so as to ensure that the chip has a better effect in the process of capturing the magnetic beads 70.
The thicknesses of the titanium seed layer 30 and the copper seed layer 40 are 0.8 to 1.2um, so that the thickness of the chip is not too large while the soft magnetic metal layer 20 is bonded to the glass substrate 10. As a preferred scheme, the thickness of the titanium seed layer 30 and the copper seed layer 40 may be 1 um.
It should be noted that, referring to fig. 3, the length of the microfluidic channel 60 in the chip is as long as possible along the moving direction of the magnetic beads 70, so as to increase the moving distance of the magnetic beads 70 in the microfluidic channel 60 as much as possible, thereby ensuring that more magnetic beads 70 are captured when an external magnetic field is applied, and completing the separation and purification of the substances in the nucleic acid extraction experiment. In one embodiment, the microfluidic channel 60 may be formed by combining a plurality of end-to-end sub-channels along the length direction of the glass substrate 10, and the form of the microfluidic channel 60 greatly increases the moving path of the magnetic beads 70, thereby further increasing the capture rate of the magnetic beads 70.
Further, the height of the microfluidic channel 60 should be determined according to the diameter of the magnetic bead 70 to be captured, and when the diameter of the magnetic bead 70 is larger, the height of the microfluidic channel 60 is correspondingly higher, so that the magnetic bead 70 can move smoothly in the microfluidic channel 60. Wherein the height of the microfluidic channel 60 is specifically set in the range of 0.5-3 mm.
In one embodiment, the height of the soft magnetic metal layer 20 in the chip can be set to 50um, so as to ensure that the soft magnetic metal layer 20 has enough magnetic force on the magnetic beads 70 when a corresponding external magnetic field is applied, so that the magnetic beads 70 are adsorbed at the bottom of the microfluidic channel 60.
It should be noted that the material of the soft magnetic metal layer 20 in the chip in the present application may be one of iron-nickel alloy, pure iron and low carbon steel, iron-silicon alloy, iron-aluminum alloy, soft magnetic iron oxide ti3, and amorphous soft magnetic alloy, and the shape of the soft magnetic metal layer 20 may be set according to different requirements, so that the chip has high universality.
Based on the same inventive concept, the present application may also provide a method for manufacturing a chip, which may refer to fig. 6 and specifically includes the following steps:
s100: forming a soft magnetic metal layer 20 on the glass substrate 10;
s200: forming a microfluidic channel 60;
s300: the microfluidic channel 60 is bonded to the glass substrate 10 such that the soft magnetic metal layer 20 is located between the glass substrate 10 and the microfluidic channel 60.
By forming the soft magnetic metal layer 20 inside the chip, the magnetic field strength of the soft magnetic metal layer 20 can be increased when an external magnetic field is applied, and the capture rate of the magnetic beads 70 can be increased. In addition, the chip prepared by the method can reduce the dependence on an external magnetic field and improve the adaptability of the chip and equipment.
Specifically, in the above step S100, as shown in fig. 7, forming the soft magnetic metal layer 20 on the glass substrate 10 may include the steps of:
s101: coating a positive photoresist on the glass substrate 10;
s102: developing and exposing the positive photoresist by using a patterning process to obtain a recessed portion, and filling a soft magnetic metal material in the recessed portion to form a soft magnetic metal layer 20;
s103: and removing the residual positive photoresist.
In step S101, the glass substrate 10 needs to be strictly cleaned between the application of the positive photoresist on the glass substrate 10, and the thickness of the glass substrate 10 may be selected to be about 0.5 mm.
Specifically, after the completion of the above step S103, i.e., after the removal of the remaining positive photoresist, a silicon oxide layer may be further deposited on the formed soft magnetic metal layer 20 to form an insulating layer 50 to protect the soft magnetic metal layer 20, and the deposition of the silicon oxide layer may be performed by a vapor deposition method.
Further, before the above step S101, i.e., before the positive photoresist is coated on the glass substrate 10, the titanium seed layer 30 and the copper seed layer 40 may be sequentially sputtered on the glass substrate 10, and when the remaining positive photoresist is removed, the titanium seed layer 30 and the copper seed layer 40 corresponding to the positive photoresist are simultaneously removed in step S103. The titanium seed layer 30 and the copper seed layer 40 are formed in the above manner, so that the formed soft magnetic metal layer 20 is better attached to the glass substrate 10.
Specifically, in the step S200, as shown in fig. 8, the forming of the microfluidic channel 60 may specifically include the following steps:
s201: coating SU-8 photoresist on a silicon wafer;
s202: developing and exposing the SU-8 photoresist by using a composition process so that the SU-8 photoresist is in the shape of the microfluidic channel 60;
s203: pouring a PDMS (polydimethylsiloxane) 80 layer on the SU-8 photoresist;
s204: and removing the SU-photoresist.
It should be noted that in the step S201, before the SU-8 photoresist is coated on the silicon wafer, the silicon wafer needs to be cleaned.
In step S300, before bonding the microfluidic channel to the glass substrate, the formed PDMS chip (i.e., the formed microfluidic channel) needs to be cleaned, and the surface needs to be subjected to oxygen plasma treatment.
It will be apparent to those skilled in the art that various changes and modifications may be made in the embodiments of the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.
Claims (13)
1. A chip, comprising:
the glass substrate, the soft magnetic metal layer and the microfluidic channel are bonded with the glass substrate, so that the soft magnetic metal layer is located between the glass substrate and the microfluidic channel.
2. The chip of claim 1, further comprising an insulating layer between the soft magnetic metal layer and the microfluidic channel.
3. The chip according to claim 1, further comprising a titanium seed layer and a copper seed layer located between the soft magnetic metal layer and the glass substrate, sequentially arranged in a direction from the glass substrate to the soft magnetic metal layer, and in one-to-one correspondence with the soft magnetic metal layer.
4. The chip of claim 3, wherein the titanium seed layer has a thickness of 0.8-1.2um, and the copper seed layer has a thickness of 0.8-1.2 um.
5. The chip of claim 1, wherein the microfluidic channel comprises a plurality of S-shaped sub-channels connected end to end in sequence along the length of the glass substrate.
6. The chip of claim 5, wherein the height of the microfluidic channel is 0.5-3 mm.
7. The chip of claim 1, wherein the thickness of the soft magnetic metal layer is 50 um.
8. The chip according to any one of claims 1 to 7, wherein the material of the soft magnetic metal layer is one of iron-nickel alloy, pure iron and low carbon steel, iron-silicon alloy, iron-aluminum alloy, soft magnetic iron oxide ti3, and amorphous soft magnetic alloy.
9. A method of manufacturing a chip according to any one of claims 1 to 8, comprising:
forming a soft magnetic metal layer on a glass substrate;
forming a microfluidic channel;
bonding the microfluidic channel to the glass substrate such that the soft magnetic metal layer is located between the glass substrate and the microfluidic channel.
10. The production method according to claim 9, wherein the forming a soft magnetic metal layer on a glass substrate comprises:
coating positive photoresist on a glass substrate;
developing and exposing the positive photoresist by using a patterning process to obtain a depressed part, and filling a soft magnetic metal material in the depressed part to form a soft magnetic metal layer;
and removing the residual positive photoresist.
11. The method of claim 10, wherein after removing the remaining positive photoresist, further comprising depositing a silicon oxide layer on the soft magnetic metal layer to form an insulating layer.
12. The method of claim 10, wherein before the applying the positive photoresist on the glass substrate, further comprising: sputtering a titanium seed layer and a copper seed layer on the glass substrate in sequence;
and when the residual positive photoresist is removed, simultaneously removing the titanium seed layer and the copper seed layer corresponding to the positive photoresist.
13. The method of manufacturing of claim 9, wherein the forming a microfluidic channel comprises:
coating SU-8 photoresist on a silicon wafer;
developing and exposing the SU-8 photoresist by using a composition process so that the SU-8 photoresist is in the shape of a microfluidic channel;
pouring a PDMS layer on the SU-8 photoresist;
and removing the SU-8 photoresist.
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| CN202011167194.6A CN114471752A (en) | 2020-10-27 | 2020-10-27 | Chip and preparation method thereof |
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| CN202011167194.6A CN114471752A (en) | 2020-10-27 | 2020-10-27 | Chip and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116121063A (en) * | 2022-12-30 | 2023-05-16 | 山东大学 | Biochip for realizing magnetic field regulation and temperature monitoring and preparation method thereof |
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| WO2019010787A1 (en) * | 2017-07-12 | 2019-01-17 | 华讯方舟科技有限公司 | Blood cell capture chip and method |
| CN109746064A (en) * | 2019-01-28 | 2019-05-14 | 武汉纺织大学 | A kind of gradient magnetic micro-fluidic chip |
| CN110079457A (en) * | 2019-06-04 | 2019-08-02 | 苏州大学 | Micro-fluidic chip and excretion body extracting method |
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2020
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| CN101879467A (en) * | 2010-06-04 | 2010-11-10 | 武汉大学 | Micro-fluidic chip for micro-magnetic field control and manufacturing method thereof |
| US20160116402A1 (en) * | 2013-07-25 | 2016-04-28 | Suzhou Institute Of Nano-Tech And Nano-Bionics Of Chinese Academy Of Science | High-Sensitivity Terahertz Micro-Fluidic Channel Sensor and Preparation Method Thereof |
| CN103869060A (en) * | 2014-03-07 | 2014-06-18 | 复旦大学附属中山医院 | Circulating tumor stem cell detection kit based on magnetic beads and microfluidic chip |
| WO2019010787A1 (en) * | 2017-07-12 | 2019-01-17 | 华讯方舟科技有限公司 | Blood cell capture chip and method |
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| CN116121063A (en) * | 2022-12-30 | 2023-05-16 | 山东大学 | Biochip for realizing magnetic field regulation and temperature monitoring and preparation method thereof |
| CN116121063B (en) * | 2022-12-30 | 2023-08-04 | 山东大学 | Biochip for realizing magnetic field regulation and temperature monitoring and preparation method thereof |
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