WO2013097262A1

WO2013097262A1 - Highly parallel microfluidic chip applicable in fabricating nanoparticles

Info

Publication number: WO2013097262A1
Application number: PCT/CN2012/000330
Authority: WO
Inventors: 陈颖; 罗春雄
Original assignee: 北京瑞斯诺生物医药技术有限公司
Priority date: 2011-12-30
Filing date: 2012-03-16
Publication date: 2013-07-04
Also published as: CN102527453A; CN102527453B

Abstract

A microfluidic chip consisting of a PDMS top layer, a PDMS intermediate layer, a PDMS bottom layer, and a glass substrate. The PDMS top layer, the PDMS intermediate layer, and the PDMS bottom layer are all provided with a groove at the front-middle portion of their respective lower surfaces. When the PDMS top layer, the PDMS intermediate layer, the PDMS bottom layer, and the glass substrate are sequentially aligned and bonded, three layers of flow paths, namely a top layer flow path, an intermediate layer flow path, and a bottom layer flow path, are formed respectively by the groove of the PDMS top layer and the PDMS intermediate layer, by the groove of the PDMS intermediate layer and the PDMS bottom layer, and by the groove of the PDMS bottom layer and the glass substrate. Three fluid inlets respectively in communication with the top layer flow path, the intermediate layer flow path, and the bottom layer flow path are sequentially arranged on the upper surface of the PDMS top layer on the basis of the positional order of the three layers of flow paths, namely the top layer flow path, the intermediate layer flow path, and the bottom layer flow path, while 10 to 1000 fluid outlets are arranged at the end of each layer of flow path. The present invention employs a scheme of multiple paths in parallel and fluid envelopment at the fluid outlets to achieve uniformity control of a product, and to prevent the product from being clogged within the flow paths.

Description

High parallel microchannel chip for nanoparticle preparation

The invention relates to a high parallel micro-flow channel chip applied to the preparation of nanoparticles, belonging to the direction of nano-pharmaceutical. Background technique

A method for synthesizing polymer nanoparticles of several tens to several hundreds of nanometers in size has been reported in the prior art. The main method is to use a microfluidic method to sandwich an organic phase of a polymer molecule having a hydrophobic group in water. Quite quite.

In the prior art body mixing method, there is a disadvantage that mixing inconsistency is likely to occur, resulting in poor uniformity of materials. Reference material 1 "Microfluidic Platform for Controlled Synthesis of Polymeric Nanoparticles", reference 2 "Micronization of Size-Tunable Polymeric Nanoparticles Enabled by 3D Hydrodynamic Flow Focusing in Single-Layer Microchannels" uses the polymer material PDMS ( Polydimethylsiloxane) The single flow channel is prepared, and the mixing of the body is reduced from a hundred micron to a micron scale. The characteristic diffusion reaction time without stirring can be smaller than the characteristic time of the nanoparticle formation, and the reaction is more uniform. As shown in Fig. 1 and Fig. 2, due to the laminar flow, the organic phase is sandwiched by the aqueous phase to a thickness of about several micrometers, and the organic solvent used is an infinitely miscible liquid with water. When the organic solvent diffuses into the aqueous phase, it dissolves. The polymer molecules in the organic solvent will self-assemble into particles of several tens of nanometers, the inside is a hydrophobic group and the outside is a hydrophilic group, the average size is smaller than the body mixing, and the uniformity is better than the body mixing, but Into the hydraulic pressure limit, the output can only reach a speed of about 20ul / h, and there is also a polymer material PDMS surface is easy to adsorb molecules, resulting in flow channel blockage. As shown in Fig. 3 and Fig. 4, PLGA polylactic acid-glycolic acid copolymer-PEG polyethylene glycol: improved 3D flow channel can avoid surface adsorption, but it still does not improve in yield, and needs more The fluid control of the road to complete the single-channel output, the product is difficult to obtain in high yield, and the industrial application prospect is poor. Summary of the invention

The technical solution of the present invention. .

A microchannel chip for nanoparticle preparation.

The micro flow channel chip, as shown in FIG. 5, FIG. 6, and FIG. 7, is composed of a PDMS top layer, a PDMS intermediate layer, a PDMS bottom layer, and a glass back sheet;

Wherein, a groove is formed in the front part of the PDMS top layer, the PDMS intermediate layer and the lower surface of the PDMS bottom layer. When the PDMS top layer, the PDMS intermediate layer, the PDMS bottom layer and the glass back sheet are sequentially aligned, the groove of the top layer of the PDMS is PDMS middle layer, The groove of the intermediate layer of the PDMS and the bottom layer of the PDMS, the groove of the bottom layer of the PDMS and the glass backsheet respectively form a top flow channel, an intermediate layer flow channel, and a bottom flow channel three-layer flow channel, and the three-layer flow channels are not connected to each other and are parallel to each other;

Wherein, on the upper surface of the top layer of the PDMS, three inlet ports communicating with each other are sequentially arranged according to the position of the top flow channel, the middle layer flow channel, and the bottom flow channel; the number of the flow channels at each end is 10 to 1000 Liquid outlet.

Wherein, the liquid inlets are perpendicular to each of the flow channels and are parallel to each other; the flow path between the liquid inlet and the liquid outlet of each layer is a tree structure, and is divided into two branch flows by one flow channel. The road is divided into three branch flow passages, which are equally divided into six branch flow passages, and are divided into 10-1000 branch flow passages as the liquid outlets.

Wherein, the liquid inlet line width is 100-400 um, the intermediate layer flow channel outlet opening width is 5-20 micrometers, the height is 5-15 micrometers, the line period is 100 micrometers; the top flow channel outlet port and the bottom layer flow channel The outlet has a width of 10-40 microns, a height of 5-20 microns, and a line period of 50 microns.

Wherein, the middle layer flow channel is an organic phase flow channel, and the top layer flow channel and the bottom layer flow channel are water phase flow channels. The organic phase is a water-miscible organic solvent and a water-insoluble solute, an organic solvent such as methanol, ethanol, isopropanol, butanol, acetone, acetonitrile, etc., the solute is a biodegradable polymer and is encapsulated Hydrophobic drugs, polymers such as: polylactic acid (PLA), polylactic acid-glycolic acid copolymer (PLGA), polyethylene glycol (PEG), polyethylene glycol-polylactic acid copolymer (PLA-PEG, PLA- PEG-PLA, PEG-PL-PEG), polyethylene glycol-polylactic acid-glycolic acid copolymer (PLGA-PEG, PLGA-PEG-PLGA, PEG-PLGA-PEG), polycaprolactone (PCL), Polyethylene glycol-polycaprolactone copolymer (PCL-PEG' PCL - PEG-PCL), polyglyconate, polyanhydrides, polyorthoesters, polydioxane Polydioxanone, polyalkylcyanoacrylates, and the like.

Among them, the intermediate layer flow channel mold is prepared by using the photoresist SU83005 model, and the top flow channel mold and the bottom flow channel mold are prepared by using the photoresist SU83025 model.

Wherein, the PDMS is prepared by mixing the monomer A and the crosslinking agent B in proportion; the PDMS ratio of the PDMS top layer is 5-20:1 (mass ratio), and the PDMS intermediate layer adopts a PDMS ratio of 5 -20 : 1 (mass ratio), PDMS bottom layer adopts PDMS ratio of 5-20: 1 (mass ratio); PDMS item layer, PDMS middle layer and PDMS bottom layer adopt different PDMS ratios.

The PDMS material can also be made of plexiglass (polymethyl methacrylate P let A) or vinyl polymer, vinyl polymer selected polystyrene PS, polyethylene PE, polyvinyl chloride PVC, polydichloroethylene PVDC Other polymer materials are replaced. .

A preparation process of a three-layer parallel microchannel chip, comprising the following steps:

Step 1: Photolithography process 1) Intermediate layer flow channel mold preparation process: prepared by using photoresist SU83005, the intermediate layer flow channel outlet opening width is 5-20 microns, the height is 5-15 microns, and the line period is 100 microns;

2) Preparation process of the top and bottom flow channel molds: Prepared by photoresist SU83025, the top flow channel outlet and the bottom flow channel outlet have a width of 10-40 microns, a height of 5-20 microns, and a line cycle of 50. Micron.

Step 2: Soft injection molding process

1) The above layers use PDMS with different ratios, and the PDMS ratio is between 5-20:1;

2) Injecting PDMS with a thickness of 20-40 microns through the silicone machine on the underlying runner mold, the corresponding speed of the silicone machine is 5000 to 2000 rpm;

3) Injecting 10-30 micron PDMS into the intermediate layer runner mold by a silicone machine, the corresponding speed of the silicone machine is 6000 to 3000 rpm;

4) Pour the PDMS with a thickness of 6mm-2cm on the top flow channel mold;

5) The above three-layer flow path molds were placed in a 75 ° C oven for 30 minutes.

Step 3: Alignment and bonding

1. After cutting the top layer of PDMS, punch the hole, treat the surface by plasma, place it on the cured intermediate layer PDMS mold, and bake in the oven at 75 °C for 30 minutes to complete the adhesion of the PDMS top layer and the PDMS intermediate layer. ;

2. Similarly, the two layers of PDMS that have been bonded are cut from the mold, and after the outlet of the intermediate layer is opened, the surface is treated by plasma, and then placed on the cured bottom layer PDMS mold, 75 degrees Celsius in the oven. Bake overnight to complete bonding to the underlying PDMS.

3. Cut the bonded three-layer PDMS, open the bottom outlet, and bond it to the glass backsheet by plasma treatment to bond the final chip. For the purposes of the present invention, the first important is product homogeneity. The thickness of the chip PDMS film is determined at the same flow rate ratio; the second is the production speed, compared with references 1, 2, the production speed has been 10-50. With a multiple increase, the number of runners can partially limit the maximum flow rate, ie the maximum production speed.

The technical effects of the invention: 1. Using the emerging MEMS (indicating Chinese meaning) and the soft lithography process, the micron-sized polymer flow channel can be conveniently prepared, and the polymer material polydimethylsiloxane (PDMS) material can be prepared. The low cost can greatly reduce the manufacturing cost of the device. 2. The chemical reaction carried out in the microchannel, the molecular precipitation, the comparison of the conventional sample mixing and mixing reaction, due to the direct decrease of the size, the diffusion into a square relationship, which can speed up the production of the product and better realize the product. Uniformity control. 3, the use of micro-scale liquid coating can shorten the mixing reaction time, let the production The material is more uniform, but recent prior art requires complex fluid control to complete sample preparation for a single channel, requiring high expertise and extremely low product yield. 4. The new device chip adopts the combination of micro-flow and body mixing, and the characteristic time of mixing is obviously smaller than that of body mixing, reducing the injection channel to micron size, and adopting high flow rate and high parallel channel to achieve high Sample uniformity control under yield.

The invention adopts multi-channel parallel and the way of the liquid outlet reaction to achieve high output, and adopts the method of controlling the size of the liquid outlet and the liquid coating of the liquid outlet to realize the uniformity control of the product. Since the reaction is carried out outside the flow channel, the product is avoided. The clogging in the flow channel is a typical application that combines the advantages of conventional bulk mixing and microfluidic systems. DRAWINGS

Figure 1 is a schematic view showing uniform nanoparticles obtained by laminar flow fast diffusion mixing;

Figure 2 is a schematic diagram of a 2D microfluidic device;

Figure 3 is a schematic diagram of an improved 3D generation nanoparticle microfluidic device;

Figure 4 is a quasi-3D hybrid mode in an improved microchannel;

Figure 5 is a plan view of the liquid outlet of each layer;

Figure 6 is a side view of the chip;

Figure 7 is a perspective view of the chip;

Figure 8 is a photograph of the inlet port;

Figure 9 is a picture of the liquid outlet under the microscope;

Figure 10 is a diagram showing the flow of the outlet laminar flow with the fluorescent molecule in place of the outlet of the intermediate layer (top view micrograph);

Figure 11 is a photo of the experiment;

Figure 12 is a comparison photograph of the products obtained by the present invention and the prior art;

Figure 13 is a particle size distribution: dynamic light scattering results of different flow rates of the same chip;

Figure 14 is a graph showing the results of changing the line width of the water channel (25 micron chip 1 to 40 micron chip 2);

Figure 15 is a comparison of the product and the body of the chip with varying layer thickness;

Figure 16 is a comparison of the product and body mixture produced by the 3D, 2D microfluidic chip in the supplementary material of Reference 2. detailed description

The invention will now be further elucidated with reference to the accompanying drawings. Example 1 A three-layer parallel microchannel chip 1 was prepared: Step 1: Photolithography process

1) Intermediate layer flow path mold preparation: Using photoresist SU83005, a 5-10 micron photoresist (rotation speed 3000 rpm-2000 rpm, time 30 s) is coated on the silicon wafer by a silicone machine, and baked at 65 degrees Celsius. 1 minute, 95 degrees Celsius baking, 5 minutes before baking, exposed to the exposure machine, the mask used is the negative layer of the designed intermediate layer pattern (Fig. 5), the exposure time is 50 seconds, and the light intensity is 20mw/ _C m ² _{; After} exposure, the sample was placed at 65 ° C for 1 minute, and after 95 ° C for 5 minutes; after development by a developing solution, an intermediate layer mold was obtained.

2) Preparation of the top and bottom runner molds: 光刻Using the photoresist SU83025, the procedure is the same as above, the pre-baking conditions are 65 degrees Celsius for 1 minute, 95 degrees Celsius for 10 minutes, the corresponding mask is used, the exposure time is 90 seconds, and the light intensity is 20mw. /cm2, post-baking conditions are 65 degrees Celsius for 1 minute, 95 degrees Celsius for 10 minutes.

As shown in Figure 5, Figure 6, and Figure 7, each layer of the pattern is an entrance divided into hundreds of outlets through a tree structure, the width of the entrance is 300um, divided into 150 microns, and then to 100 microns. Strips, 50 micron 6 strips, 50 micron 12 strips, to exit 10-1000 flow channels. The top of the exit runner width is 25 microns wide with a period of 50 microns and the middle layer is 5 microns wide with a 100 micron period.

Step 2: Soft injection molding process,

1) PDMS with different ratios is used. The PDMS ratio of the top layer of the PDMS is 15:1, the PDMS ratio of the PDMS middle layer is 10:1, and the PDMS bottom layer is 8:1.

2) Injecting a 30-micron-thick PDMS onto the underlying runner mold by a silicone machine, the corresponding speed of the silicone machine is 2000 rpm;

3) Inject 20 micron PDMS into the middle layer flow path mold through a rubberizing machine, and the corresponding speed of the rubberizing machine is 3000 rpm;

4) Pour the PDMS with a thickness of 6mm on the top flow channel mold;

5) The above three layers of PDMS covered runner molds were placed in a 75 ° C oven for 30 minutes.

Step 3: Alignment and bonding

1) After cutting the top layer of PDMS, punch the hole, treat the surface by plasma, place it on the cured intermediate layer TOMS mold, and bake in the oven at 75 ° C for 30 minutes to complete the adhesion of the PDMS top layer and the PDMS intermediate layer. ;

2) Similarly, the two layers of PDMS that have been bonded are cut from the mold, and after the outlet of the intermediate layer is opened, the surface is treated by plasma, and then placed on the cured bottom layer PDMS mold, 75 degrees Celsius in the oven. Bake overnight to complete bonding to the underlying PDMS. ' '

3) cutting the bonded three-layer PDMS to open the bottom layer of the outlet and bonding it to the glass backsheet by plasma treatment to form a final core sheet; The resulting chip 1 has a line width of 300 μm, a PDMS intermediate layer with 100 liquid outlets, and a PDMS top end with 240 outlets. The PDMS intermediate channel has a 5 μm width. 5 micron, line period is 100 microns; PDMS top flow channel outlet and PDMS bottom channel outlet width is 25 microns, height is 15 microns, line period is 50 microns; PDMS bottom layer thickness is 30 microns, PDMS middle The layer thickness is 20 microns. Example 2 Preparation of a three-layer parallel microchannel chip 2:

By the method of the first embodiment, the inlet width of the three-layer parallel microchannel chip is 300 nm, the outlet of the intermediate layer is provided with 50 liquid outlets, and the end of the top layer is provided with 120 liquid outlets. The outlet of the intermediate flow channel has a width of 20 μm, a height of 15 μm, and a line period of 100 μm. The outlet of the top flow channel and the bottom channel has a width of 40 μm, a height of 20 μm, and a line period of 50. Micron; PDMS underlayer thickness is 40 microns, PDMS interlayer thickness is 30 microns. Example 3 Preparation of a three-layer parallel microchannel chip 3:

The method of Example 1 was used, but the PDMS was replaced by a plexiglass crucible, and the crucible was heated and liquefied, which was not the same cross-linking curing reaction as PDMS; the PDMS of the first step in the second step of Example 1 could be changed to heat melting at 180 ° C. After PMMA; 5) The resulting three-layer PMMA-covered runner mold was cooled at room temperature; in step three, the plasma bond was changed to heat-bond at 140 °C.

The three-layer parallel micro-channel chip inlet has a line width of 100 μm, the middle layer flow channel has 450 liquid outlets, and the top and bottom layers have 1000 liquid outlets. The outlet of the intermediate flow channel has a width of 10 μm, a height of 10 μm, and a line period of 100 μm; the outlet of the top flow channel and the bottom channel has a width of 10 μm, a height of 5 μm, and a line period of 50. Micron; PMMA underlayer thickness is 35 microns, PMMA interlayer thickness is 25 microns. Example 4 Preparation of a three-layer parallel microchannel chip 4:

By the method of the first embodiment, the inlet width of the three-layer parallel microchannel chip is 400 _um , the outlet of the middle layer flow channel is provided with 10 liquid outlets, and the end of the top layer is provided with 20 liquid outlets. The outlet of the intermediate flow channel has a width of 20 μm, a height of 3⁄4 is 15 μm, and a line period of 100 μm; the outlet of the top flow channel and the bottom channel has a width of 40 μm and a height of 20 μm. It is 50 microns; the PDMS underlayer has a thickness of 25 microns and the PDMS interlayer has a thickness of 15 microns. Chip result test:

1, liquid control The singularity of the singularity of the singularity of the singularity of the singularity of the liquid. After a minute, the intermediate layer flow channel inlet is passed through a PEG-PLGA/ACN solution with a typical concentration of 50 mg/ml. The typical polymer molecular weight is PEG _5k - PLGA _55k , with a typical push rate of 100 ul/h to 1 ml/h. The sample was collected after one hour of pushing and pure water was added to a volume of 10 ml.

The body mixing conditions were such that 200 ul of a 50 mg/ml PEG-PLGA/ACN solution was added to 10 ml of the liquid.

2. Identification of samples

As shown in Figure 12, from left to right, the mixture product A, 200 ul volume, 50 mg/ml concentration of PLGA _55k - PEG _5k / ACN was added to 10 ml of pure water; chip product 8, C, D, 50 mg The concentration of the particle concentration is 1. 5ml / h, reflecting the particle concentration of the PLGA _55k - PEG _5k / ACN solution, the rate of 200ul / h, 500ul / h, lral / h, the total liquid flow time is 1 hour, the water phase flow rate is 1. 5ml / h, reflecting the particle concentration That is, lmg/ml, 2. 5mg/mL 5mg/ml, the particle size is measured as shown in Figure 13, Figure 14, the product B, C, D is about 150-160nm, the peak is narrow, and the body mixing result product A The particle size is extremely heterogeneous and many micron-sized particles are produced. Through the test, the following conclusions can be drawn:

1) High throughput. As shown in Fig. 5, Fig. 6, Fig. 7, and Fig. 9, the flow path of the tree structure is branched into a plurality of flow paths by one flow path, which can greatly increase the flow rate of the chip through which the liquid passes.

2) Quick mixing effect. Control Reference 1 and Reference 2, Figure 10 is a picture under the microscope, from which it can be seen that the organic phase and the aqueous phase are mixed at the liquid outlet.

3) High product uniformity. It can be seen from Fig. 13 that the sample homogeneity result of the chip is obviously better than that of the body mixture, and the body mixing obviously has large-sized particles, and the flow rate is increased to increase the yield, and the product properties are not significantly changed (organic phase 200 ul/h and lOOOOul/h, the molecule is PLGA). _55k -PEG _5k, each of the aqueous phase is 1. 5ml / h). The production rate is ten times higher than that of the existing literature ("Microfluidic Platform for Controlled Synthesis of Polymeric Nanoparticles", "Synthesis of Size-Tunable Polymeric Nanoparticles Enabled by 3D Hydrodynamic Flow Focusing in Single-Layer Microchannels"), and the particle size is uniform. The results were close to those reported in the literature, and the product was close at a flow rate of 200 ul/h to 1 ml/h, indicating that the flow rate control accuracy was low and suitable for industrial production.

Theoretical and test results for different chips: As shown in the figure, the flow rate is 500ul/h, the molecule is PLGA _55k - PEG _5k , and the line width of the water channel can get better coating effect. The applicant tested two types of water flow. The width of the chip: 25 μm width and 40 μm width, the product was found to be substantially unchanged.

As shown in Fig. 15, PLGA _55k -PEG _5k 50mg/ml, the pushing speed is 500ul/h, chip 1 is the bottom film thickness 30um, in the middle The layer thickness is 20 ura, the chip 2 has an underlying film thickness of 40 μm, and the intermediate layer film has a thickness of 30 μm. The test result chip products are superior to the body mixed product. Each channel of the organic phase should be far enough apart to satisfy the surrounding area of each organic phase and be surrounded by water. Theoretically, the thinner the liquid phase and the organic phase liquid layer, the thinner the degree of thickness directly affects the diffusion time at the same flow rate. The thinner the product, the better the product uniformity, and in the case of the same flow rate ratio, the chip PDMS bottom layer and The thickness of the middle layer directly affects the thickness of the liquid layer. We tested two types of chips, as shown in Figure 15. The chip 1 has a film thickness of 30 μm for the bottom layer and 20 μm for the middle layer. The same flow rate is used, and the result is slightly better than the bottom layer. 40 micron, the middle layer is 30 micron chip 2, but due to the PDMS silicone curve, the lower limit of PDMS film thickness is 15 microns, and the thinner, the worse the stability of the chip, so we choose the optimal condition. The bottom layer is 30 microns and the intermediate layer is 20 microns thick.

Figure 16 is a supplement from the reference article "Synthesis of Size-Tunable Polymeric Nanoparticles Enabled by 3D Hydrodynamic Flow Focusing in Single-Layer Microchannels", which indicates the product prepared by bulk mixing. The higher the concentration of PLGA-PEG, the larger the product. And the heterogeneity, the product of the present invention is close to the 3D, 2D product size and uniformity at the same concentration and close molecular weight, but the flow rate control range is from 200 ul / h to 1000 ul / h, the product is not Too much change can explain our conclusions above.

4) When the chip provided by the invention is from 200 ul/h to lOOOul/h, the products are close to the literature results, and the comparative literature needs to strictly limit the flow rate, and the PLGA-PEG/ACN flow rate is much lower than the case of the present invention. The ultimate push rate of the chip of the present invention can be 15 ml/h in the aqueous phase and 5 ml/h in the organic phase. Under this condition, the product size of the same concentration and molecular weight does not change much. It has also been proved that the chip has low flow rate control precision requirements and large output, and is more suitable for industrial production than the existing microfluidic literature.

Compared with the mixed body, the product obtained by the chip of this patent has good uniformity; compared with Reference 1 and Reference 2, only 3 channels are needed, the product speed is more than 10 times faster, and the product uniformity index is close to it. The above analysis can be concluded. Conclusion Our chips and related technical solutions can achieve high yield and uniform nanoparticle preparation, and its low flow rate control makes it suitable for industrial production.

Claims

Claim

A microchannel chip characterized by comprising a PDMS top layer, a PDMS intermediate layer, a PDMS underlayer and a glass substrate.

2. The microchannel chip according to claim 1, wherein a groove is provided in the front portion of the PDMS top layer, the PDMS intermediate layer and the lower surface of the PDMS underlayer; when the PDMS top layer, the PDMS intermediate layer, the PDMS bottom layer and the glass When the negative film is sequentially aligned, the groove of the top layer of the PDMS and the PDMS intermediate layer, the groove of the PDMS intermediate layer and the PDMS bottom layer, the groove of the PDMS bottom layer and the glass back sheet respectively form a top flow channel, an intermediate layer flow channel, and a bottom flow channel. The three-layer flow channel and the three-layer flow channel are not connected to each other and are parallel to each other.

3. The micro-channel chip according to claim 2, wherein, on the upper surface of the top layer of the PDMS, three adjacent channels are sequentially arranged according to the position of the top flow channel, the middle layer flow channel, and the bottom layer flow channel Liquid port; a liquid outlet is provided at the end of each layer of the flow channel.

4. The microchannel chip according to claim 3, wherein the liquid inlet is perpendicular to each of the flow channels and parallel to each other; the flow path between the liquid inlet and the liquid outlet of each layer is a tree shape The structure is divided into two branch flow channels by one flow channel, and then divided into three branch flow channels, which are equally divided into six branch flow paths, and are divided into 10-1000 branch flow paths. Liquid

P.

The micro-channel chip according to claim 3 or 4, wherein the liquid inlet line has a line width of 100-400 um, and each layer is provided with 10-1000 liquid outlets; and the intermediate layer flow channel outlet opening width is 5- 20 μm, 5-15 μm twist, 100 μm line period; top channel flow port and bottom channel outlet width 10-40 μm, height 5-20 μm, line cycle 50 Micron; PDMS underlayer thickness is 20-40 microns, PDMS interlayer thickness is 10-30 microns.

6. The microchannel chip according to claim 5, wherein the liquid inlet line has a width of 300 um, and each layer is provided with 100 liquid outlets; the intermediate layer flow passage has a liquid outlet width of 5 micrometers and a height of 5 micrometers. The line period is 100 microns: the top channel outlet and the bottom channel have a width of 25 microns, a height of 15 microns, a line period of 50 microns, a PDMS bottom layer thickness of 30 microns, and a PDMS interlayer thickness of 20 microns. .

7. The microchannel chip according to claim 6, wherein the intermediate layer flow channel is an organic phase flow channel, and the top layer flow channel and the bottom layer flow channel are water phase flow channels.

The microchannel chip according to claim 7, wherein the organic phase is composed of a water-miscible organic solvent and a water-insoluble solute.

The microchannel chip according to claim 8, wherein the organic solvent is one or more selected from the group consisting of methanol, ethanol, isopropanol, butanol, acetone, and acetonitrile.

The microchannel chip according to claim 8, wherein the solute selects a biodegradable polymer and an encapsulated hydrophobic drug.

11. The microchannel chip according to claim 10, wherein the biodegradable polymer is selected from the group consisting of polylactic acid, polylactic acid-glycolic acid copolymer, polyethylene glycol, polyethylene glycol-polylactic acid copolymer, Polyethylene glycol-polylactic acid-glycolic acid copolymer, polycaprolactone, polyethylene glycol-polycaprolactone copolymer, polygluconic acid ester, polyfluorene, polyorthoester, polydioxane One or more of cycloketone, polydecyl cyanoacrylate.

The micro flow channel chip according to claim 5, wherein the three-layer flow path molds are all prepared by using a photoresist.

The microchannel chip according to claim 5, wherein the PDMS is prepared by mixing the monomer A and the crosslinker B in proportion; and the PDMS mass ratio of the PDMS top layer is 5-20: 1. The PDMS quality ratio of the PDMS intermediate layer is 5-20: 1, the PDMS bottom layer adopts the PDMS mass ratio ratio of 5-20:1; the PDMS top layer, the PDMS intermediate layer, and the PDMS bottom layer adopt the PDMS ratio. They are all different.

14. The microchannel chip according to claim 5, wherein the PDMS material is replaced with an organic glass polymethyl methacrylate P匪A or a vinyl polymer; the vinyl polymer selects polystyrene One or more of PS, polyethylene PE, polyvinyl chloride PVC, and polydichloroethylene PVDC.