CN110523449B - Preparation method of transparent ceramic micro-fluidic chip - Google Patents
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- 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
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
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- C04B35/00—Shaped ceramic products characterised by their composition; Ceramics compositions; Processing powders of inorganic compounds preparatory to the manufacturing of ceramic products
- C04B35/01—Shaped ceramic products characterised by their composition; Ceramics compositions; Processing powders of inorganic compounds preparatory to the manufacturing of ceramic products based on oxide ceramics
- C04B35/447—Shaped ceramic products characterised by their composition; Ceramics compositions; Processing powders of inorganic compounds preparatory to the manufacturing of ceramic products based on oxide ceramics based on phosphates, e.g. hydroxyapatite
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- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
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- C04B2235/00—Aspects relating to ceramic starting mixtures or sintered ceramic products
- C04B2235/02—Composition of constituents of the starting material or of secondary phases of the final product
- C04B2235/30—Constituents and secondary phases not being of a fibrous nature
- C04B2235/32—Metal oxides, mixed metal oxides, or oxide-forming salts thereof, e.g. carbonates, nitrates, (oxy)hydroxides, chlorides
- C04B2235/3205—Alkaline earth oxides or oxide forming salts thereof, e.g. beryllium oxide
- C04B2235/3208—Calcium oxide or oxide-forming salts thereof, e.g. lime
- C04B2235/3212—Calcium phosphates, e.g. hydroxyapatite
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- C04B2235/70—Aspects relating to sintered or melt-casted ceramic products
- C04B2235/96—Properties of ceramic products, e.g. mechanical properties such as strength, toughness, wear resistance
- C04B2235/9646—Optical properties
- C04B2235/9653—Translucent or transparent ceramics other than alumina
Abstract
The invention discloses a preparation method of a transparent ceramic microfluidic chip, which comprises the steps of firstly, designing a substrate model with a micro-groove pattern by utilizing modeling software, printing a ceramic substrate green body by utilizing a ceramic photocuring technology, and then degreasing and sintering the substrate green body at high temperature and high pressure to realize transparentization; the transparent ceramic substrate and the PDMS sheet are bonded into a whole through the glue middle layer, a hole is punched on the PDMS sheet and connected with a conduit, and the PDMS sheet can be put into use after being introduced into microfluid. The transparent ceramic micro-fluidic chip prepared by the invention can highly simulate the bone environment, can realize high-flux drug screening, innovatively uses hydroxyapatite as the material of the micro-fluidic chip, and the good biocompatibility of the hydroxyapatite is favorable for the adhesion and growth of cells on the surface of the chip. The material is transparent by high-temperature and high-pressure sintering modes such as hot isostatic pressing and the like, and the optical property requirements of the microfluidic chip are met. The PDMS sheet is coated on the surface of the ceramic, so that cells can be cultured on the chip for a long time.
Description
Technical Field
The invention relates to the technical field of transparent ceramic preparation and microfluidic chips, in particular to a preparation method of a transparent ceramic microfluidic chip.
Background
Traditional microfluidic chip materials do not mimic bone microenvironment well. The initial microfluidics technology used the photolithography technique to build microchannel structures on silicon wafers or glass, which was time consuming and laborious. The advent of Polydimethylsiloxane (PDMS) has increased the efficiency of microfluidic technology and greatly reduced cost, but PDMS also has its limitations as an osseointegral chip. PDMS sometimes absorbs or leaches non-polar or weakly polar small molecules and is incompatible with organic solvents, requiring tedious surface modification prior to seeding the cells. Most importantly, the PDMS component is completely different from the calcium phosphate component in bone. Hydroxyapatite (HAP) is a main inorganic component of human and animal bones, can participate in vivo metabolism, has bioactivity, and is an ideal material for serving as an osseous organ chip to simulate the human bone microenvironment. The photocuring ceramic 3D printing technology has the advantages of high forming precision, complex structure and one-step forming, and solves the problem that ceramic materials are fragile and difficult to process; can be designed in a personalized way and is an ideal choice for preparing the ceramic microfluidic chip.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a preparation method of a transparent ceramic micro-fluidic chip, the prepared transparent ceramic micro-fluidic chip can highly simulate a bone environment and can realize high-flux drug screening, hydroxyapatite is innovatively used as a material of the micro-fluidic chip by the method, and the good biocompatibility of the hydroxyapatite is favorable for the adhesion and growth of cells on the surface of the chip. The material is transparent by high-temperature and high-pressure sintering modes such as hot isostatic pressing and the like, and the optical property requirements of the microfluidic chip are met. The surface of the ceramic is covered with a layer of polydimethylsiloxane cover plate (hereinafter referred to as PDMS sheet), and the good oxygen permeability of the PDMS sheet ensures that cells can be cultured on the chip for a long time.
In order to achieve the purpose, the technical scheme provided by the invention is as follows: a preparation method of a transparent ceramic microfluidic chip comprises the following steps:
1) designing a substrate part of the microfluidic chip in modeling software, wherein the surface of the substrate is provided with a micro-groove pattern, and exporting the designed substrate as an stl model;
2) leading the substrate model obtained in the step 1) into a photocuring ceramic printer, adopting a photocuring ceramic printing technology, printing layer by layer, and forming in one step, wherein the used printing material is hydroxyapatite to obtain a ceramic substrate green body of the microfluidic chip;
3) placing the ceramic substrate green body obtained in the step 2) into a high-temperature furnace for slow degreasing sintering, and then sintering at high temperature and high pressure to obtain a transparent ceramic substrate;
4) mixing a dimethoxysiloxane main agent and a hardening agent in a mass ratio of 10: 1, uniformly mixing in proportion to obtain a mixed solution, floating bubbles in the mixed solution to the surface and breaking the bubbles by using a vacuumizing mode, and then putting the mixed solution into an oven for heating and curing for a period of time to obtain a polydimethylsiloxane cover plate, namely a PDMS sheet for short;
5) cutting the PDMS sheet obtained in the step 4) according to the shape of the transparent ceramic substrate and the micro-groove pattern, punching holes at corresponding positions, wherein the diameter of the punched holes is consistent with the outer diameter of a conduit needing to be externally connected, and obtaining the punched PDMS sheet;
6) and (3) bonding the PDMS sheet obtained in the step (5) with the transparent ceramic substrate obtained in the step (3) through glue, and then thermally curing to obtain a target product, thus completing the manufacture of the transparent ceramic microfluidic chip.
Further, inserting the transparent ceramic microfluidic chip obtained in the step 6) into an external conduit, and introducing the conduit into the microfluidic chip, so that the transparent ceramic microfluidic chip can be put into use.
Further, the method for manufacturing the ceramic substrate green body adopts Stereo Light curing and forming (SLA) or Digital Light Processing (DLP).
Further, the method for making the ceramic substrate transparent is one or a combination of a hot isostatic pressing method, an atmosphere and vacuum sintering method and a discharge plasma sintering method.
Further, the catheter is made of one or a combination of PTFE, PE and PEEK.
Further, the outer diameter of the conduit is 0.5 mm-2 mm.
Further, the glue is liquid dimethyl siloxane diluted by dichloromethane, and the dilution ratio is 30% -100%.
Further, in the step 4), the curing temperature of the PDMS sheet is 65-85 ℃, and the curing time is 1-4 h.
Compared with the prior art, the invention has the following advantages and beneficial effects:
the transparent ceramic micro-fluidic chip with highly bionic bone microenvironment, which is obtained by the method of the invention, has the advantages of high temperature resistance, acid and alkali corrosion resistance, wear resistance and the like, does not need to carry out fussy surface modification before seeding cells, can be used for realizing high-throughput drug screening, exploring the interaction between bone cells and other cells and the like, and has great practical popularization value.
Drawings
Fig. 1 is a schematic diagram of a substrate structure of a ceramic microfluidic chip.
Fig. 2 is an exploded view of a transparent ceramic microfluidic chip.
Detailed Description
The present invention will be further described with reference to the following specific examples.
Example 1
Firstly, a Christmas tree type microfluidic chip is drawn by utilizing rhino software, the width of a channel is 200 mu m, the height of the channel is 200 mu m, four groups of parallel channels are formed, and an stl model file is exported. And secondly, printing the model by using a DLP (digital light processing) ceramic printer, wherein the used printing material is hydroxyapatite to obtain a ceramic substrate green body of the microfluidic chip, and referring to fig. 1, in the figure, 1 is a substrate, and 2 is a micro-groove pattern on the substrate. And thirdly, placing the printed ceramic substrate green body in a high-temperature furnace for degreasing and sintering, keeping the temperature at the highest temperature of 1280 ℃ for 1 hour. And then placing the ceramic substrate into a hot isostatic pressing sintering furnace (of course, the atmosphere and the vacuum sintering method or the discharge plasma sintering method can be selected, and the combination of the atmosphere and the vacuum sintering method or the discharge plasma sintering method can also be selected), and keeping the temperature at 1200 ℃ for 5h at 3 ℃/min to obtain the transparent ceramic substrate. Fourthly, mixing the dimethoxysiloxane main agent and the hardening agent according to the mass ratio of 10: 1, uniformly mixing, vacuumizing for 20min, and curing at 65 ℃ for 4h to obtain a polydimethylsiloxane cover plate (PDMS sheet for short), which is shown in figure 2, wherein 3 is the PDMS sheet. And fifthly, cutting the PDMS sheet into the size consistent with that of the ceramic substrate, and punching with the punching diameter of 1 mm. A sixth step of mixing 10: the liquid dimethyl siloxane of 1 is diluted to 30 percent, is coated on a PDMS sheet, is stuck with a transparent ceramic substrate together, is put into an oven and is cured for 2 hours at the temperature of 75 ℃. Seventhly, inserting three catheters (which can be one or a combination of more of PTFE, PE and PEEK) with the outer diameters of 0.5 mm-2 mm (preferably 1mm in the embodiment), and referring to fig. 2, wherein 4 is a catheter, and completing the manufacture of the microfluidic chip. And eighth step, verifying: the microfluidic chip is filled with cell suspension liquid, the cell density is 100 ten thousand per milliliter, after one day of culture, a cell live and dead fluorescent staining agent is filled, after half an hour of staining, the chip is placed under a fluorescent microscope for observation, the cell state is good, the number of live cells is far more than that of dead cells, and the Christmas tree type ceramic microfluidic chip meets the use requirement of cell culture.
Example 2
Firstly, drawing a Christmas tree type micro-fluidic chip by utilizing solidworks software, wherein the width of a channel is 100 mu m, the height of the channel is 100 mu m, eight groups of parallel channels are formed, and an stl model file is exported. And secondly, printing the model by using an SLA ceramic printer, wherein the used printing material is hydroxyapatite, and obtaining a ceramic substrate green body of the microfluidic chip. And thirdly, placing the printed substrate green body in a high-temperature furnace for degreasing and sintering, keeping the temperature at the highest temperature of 1280 ℃ for 1 hour. And then placing the ceramic substrate into a hot isostatic pressing sintering furnace (of course, the atmosphere and the vacuum sintering method or the discharge plasma sintering method can be selected, and the combination of the atmosphere and the vacuum sintering method or the discharge plasma sintering method can also be selected), and keeping the temperature at 5 ℃/min and 1200 ℃ for 3h to obtain the transparent ceramic substrate. Fourthly, mixing the dimethoxysiloxane main agent and the hardening agent according to the mass ratio of 10: 1 proportion, and curing for 1h at 85 ℃ after vacuumizing for 20min to obtain the polydimethylsiloxane cover plate (PDMS sheet for short). And fifthly, cutting the PDMS sheet into the size consistent with that of the ceramic substrate, and punching with the diameter of 1.4 mm. A sixth step of mixing 10: the liquid dimethyl siloxane of 1 is diluted to 100 percent, is coated on a pdms sheet, is stuck with a transparent ceramic substrate together, is put into an oven and is cured for 1 hour at 80 ℃. Seventhly, inserting three catheters (which can be one or a combination of PTFE, PE and PEEK) with the outer diameters of 0.5 mm-2 mm (preferably 1.4mm in the embodiment), and finishing the manufacturing of the microfluidic chip. And eighth step, verifying: the red and green fluorescent inks are respectively introduced into the external conduits on the two sides of the microfluidic chip, the microfluidic chip is placed under a laser confocal microscope for observation, the fluorescence intensity of each parallel channel is measured, and a good gradient distribution rule is displayed.
The above-mentioned embodiments are merely preferred embodiments of the present invention, and the scope of the present invention is not limited thereto, so that the changes in the shape and principle of the present invention should be covered within the protection scope of the present invention.
Claims (7)
1. A preparation method of a transparent ceramic microfluidic chip is characterized by comprising the following steps:
1) designing a substrate part of the microfluidic chip in modeling software, wherein the surface of the substrate is provided with a micro-groove pattern, and exporting the designed substrate as an stl model;
2) leading the substrate model obtained in the step 1) into a photocuring ceramic printer, adopting a photocuring ceramic printing technology, printing layer by layer, and forming in one step, wherein the used printing material is hydroxyapatite to obtain a ceramic substrate green body of the microfluidic chip;
3) placing the ceramic substrate green body obtained in the step 2) into a high-temperature furnace for slow degreasing sintering, and then sintering at high temperature and high pressure to obtain a transparent ceramic substrate;
4) mixing a dimethoxysiloxane main agent and a hardening agent in a mass ratio of 10: 1, uniformly mixing in proportion to obtain a mixed solution, floating bubbles in the mixed solution to the surface and breaking the bubbles by using a vacuumizing mode, and then putting the mixed solution into an oven for heating and curing for a period of time to obtain a polydimethylsiloxane cover plate, namely a PDMS sheet for short;
5) cutting the PDMS sheet obtained in the step 4) according to the shape of the transparent ceramic substrate and the micro-groove pattern, punching holes at corresponding positions, wherein the diameter of the punched holes is consistent with the outer diameter of a conduit needing to be externally connected, and obtaining the punched PDMS sheet;
6) bonding the PDMS sheet obtained in the step 5) with the transparent ceramic substrate obtained in the step 3) through glue, and then thermally curing to obtain a target product, wherein the transparent ceramic microfluidic chip is manufactured; the glue is liquid dimethyl siloxane diluted by dichloromethane, and the dilution ratio is 30-100%.
2. The method for preparing a transparent ceramic microfluidic chip according to claim 1, wherein the method comprises the following steps: inserting the transparent ceramic microfluidic chip obtained in the step 6) into an external conduit, and introducing the conduit into the microfluidic chip to enable the transparent ceramic microfluidic chip to be put into use.
3. The method for preparing a transparent ceramic microfluidic chip according to claim 1, wherein the method comprises the following steps: the manufacturing method of the ceramic substrate green body adopts three-dimensional photocuring forming or digital light processing.
4. The method for preparing a transparent ceramic microfluidic chip according to claim 1, wherein the method comprises the following steps: the method for making the ceramic substrate transparent is one or a combination of a hot isostatic pressing method, an atmosphere and vacuum sintering method and a discharge plasma sintering method.
5. The method for preparing a transparent ceramic microfluidic chip according to claim 1, wherein the method comprises the following steps: the catheter is made of one or a combination of PTFE, PE and PEEK.
6. The method for preparing a transparent ceramic microfluidic chip according to claim 1, wherein the method comprises the following steps: the outer diameter of the conduit is 0.5 mm-2 mm.
7. The method for preparing a transparent ceramic microfluidic chip according to claim 1, wherein the method comprises the following steps: in the step 4), the curing temperature of the PDMS sheet is 65-85 ℃, and the curing time is 1-4 h.
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| CN111685755B (en) * | 2020-05-29 | 2021-09-21 | 华南理工大学 | AgNWs-PDMS composite flexible conductive optical fiber micro-lens and preparation method thereof |
| CN113368915A (en) * | 2021-06-28 | 2021-09-10 | 上海交通大学 | Micro-fluidic chip for high-flux culture of brain organoids |
| CN115106034A (en) * | 2022-06-23 | 2022-09-27 | 之江实验室 | Visual microreactor and preparation method thereof |
| CN115193498B (en) * | 2022-07-26 | 2024-04-16 | 之江实验室 | Ceramic micro-fluidic chip and preparation method and application thereof |
| CN115055137B (en) * | 2022-08-04 | 2024-02-06 | 之江实验室 | Processing method of microreactor |
| CN115304376B (en) * | 2022-08-09 | 2023-04-18 | 之江实验室 | Transparent ceramic micro-fluidic chip and preparation method thereof |
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