CN110773244A - Micro-fluidic chip for high-throughput screening of nano-particles in cross-vascular transport and preparation method thereof - Google Patents

Abstract

A microfluidic chip for high-throughput screening of nanoparticle transvascular transport and preparation method, the microfluidic chip comprises a fluid channel layer and a glass support layer connected with the fluid channel layer, the lower end surface of the fluid channel layer is In contact with the upper end surface of the glass support layer, the fluid channel layer is provided with a fluid channel I and a fluid channel II, one end of the fluid channel I is provided with a sample inlet, and the other end of the fluid channel I is provided with a sample outlet The middle part of the fluid channel I communicates with the fluid channel II through a gap channel, and one end of the fluid channel II is provided with a seepage outlet. The microfluidic chip can be used to screen drug-loaded nanoparticles suitable for penetrating blood vessel walls, and the screening accuracy is high.

Description

Microfluidic chip for high-throughput screening of nanoparticle transvascular transport and preparation method

technical field

The invention relates to the technical field of applying microfluidic chip technology to biomedical detection and screening, in particular to a microfluidic chip for high-throughput screening of nanoparticle transvascular transport and a preparation method.

Background technique

Tumor has now become the number one killer threatening human health, and high-efficiency anti-tumor drugs have become a major demand for people's healthy life. In recent years, with the development of nanotechnology, nanoparticle-loaded drug therapy for tumors has been widely studied. Nanotechnology can not only improve the water solubility of the drug, improve the pharmacokinetic parameters of the drug in the body, but also control the release rate of the drug. However, so far, the treatment effect is still not ideal. The reason is that there are many differences between the tumor microenvironment and the normal internal environment of the human body in terms of physicochemical properties, the most notable being its characteristics of low oxygen, low pH, high osmotic pressure and high interstitial fluid pressure. It is precisely because of these characteristics that the drug cannot reach the tumor tissue in a uniform and effective concentration, resulting in insufficient local drug concentration. This heterogeneous drug distribution is the main reason for the poor therapeutic effect of drug-loaded nanoparticles. Studying how to make the nanoparticles reach the tumor to the maximum extent is the key to the treatment of tumors, so it is necessary to fully understand the transport process of nanoparticles in the human body.

The transport process of nanoparticles in the human body can be roughly divided into four stages: transport in blood vessels, penetrating the blood vessel wall into the interstitial space, transporting in the interstitial space and entering tumor cells. Among them, the transport through the vascular wall plays an important role in the entire transport process. Due to the lack of basement membrane and adhesion proteins in the tumor vascular wall, there are many holes in the vascular wall, so the nanoparticles can penetrate through these holes. Vascular wall. However, few studies have been conducted in this area, and a clear understanding of the physicochemical properties of nanoparticles and the influence of the environment in the blood vessel on their efficiency of penetrating the vessel wall is still lacking. Therefore, establishing a device to screen nanoparticles suitable for penetrating the vascular wall and suitable vascular environment is the primary task of current tumor therapy research.

Microfluidics is an emerging interdisciplinary technology that has developed rapidly in recent years. By constructing microchannels on a chip, tiny fluids can be processed or manipulated, and a series of microfabrication and micromanipulations that are difficult to accomplish by conventional methods can be realized. It can provide more accurate operations in the spatial dimension, and the scale is closer to the actual scale. Therefore, the microfluidic chip can be used as a platform for simulating the internal environment of the human body. At present, the use of microfluidic chips to screen nanoparticles suitable for penetrating the vascular wall and suitable vascular environment is still in the blank.

SUMMARY OF THE INVENTION

The technical problem to be solved by the present invention is to provide a microfluidic chip for high-throughput screening of nanoparticle transvascular transport and a preparation method for the deficiencies in the prior art. The microfluidic chip can be used for screening The drug-loaded nanoparticles suitable for penetrating the blood vessel wall have high screening accuracy and can meet the needs of modern drug-loaded nanoparticles for medical research on vascular barrier penetration.

The solution is realized by the following technical measures: a microfluidic chip for high-throughput screening of nanoparticle transvascular transport, which is characterized by comprising: a fluid channel layer and a glass support layer connected to the fluid channel layer; the The lower end surface of the fluid channel layer is in contact with the upper end surface of the glass support layer, the fluid channel layer is provided with a fluid channel I and a fluid channel II, one end of the fluid channel I is provided with a sample inlet, and the fluid channel I is provided with a sample inlet. The other end is provided with a sample outlet, the middle part of the fluid channel I is communicated with the fluid channel II through a gap channel, and one end of the fluid channel II is provided with a seepage outlet. The drug-loaded nanoparticle solution is injected into the fluid channel I from the injection port through the pressure application device. When passing through the gap channel, a part of the drug-loaded nanoparticle solution will flow out from the sample outlet through the fluid channel I, and the other part of the drug-loaded nanoparticle solution will pass through the gap. The channel enters fluid channel II and flows out from the permeate port, and the sample solution flowing out of the permeate port is collected. The above process is repeated by replacing the drug-loaded nanoparticle solution, and the drug-loaded nanoparticles that are more suitable for penetrating the tumor blood vessel wall can be screened by comparing the content of the drug-loaded nanoparticles in the collected sample solution.

The microfluidic chip is prepared by a molding method, including the following steps:

(1) Cleaning the single crystal silicon wafer: rinse with deionized water, blow dry with nitrogen after completion, and then place it on a heating table to stand;

(2) Glue coating: apply positive photoresist to the surface of the single crystal silicon wafer and spin it;

(3) Baking: put the spin-coated monocrystalline silicon wafer on the heating table and let it stand;

(4) Exposure: place the photolithography mask on the surface of the photoresist for exposure;

(5) Development: develop the exposed single crystal silicon wafer, then clean the surface of the single crystal silicon wafer with deionized water, and dry it with nitrogen;

(6) Post-baking: the single crystal silicon wafer is placed on a heating table to stand, so that the photoresist on the surface of the silicon wafer is completely cured, and a silicon-based mold is obtained;

(7) Constructing a microfluidic chip: First mix PDMS and curing agent in a volume ratio of 10:1 and stir evenly, pour it on a silicon-based mold, remove air bubbles, and place it on a heating table to allow PDMS to completely cure. , and then peel off the fully cured PDMS from the silicon-based mold to obtain the PDMS matrix, which is the fluid channel layer of the microfluidic chip;

(8) Cleaning the glass negative film: Rinse the glass negative film with absolute ethanol and deionized water in sequence, and then dry it with nitrogen;

(9) Bonding: Put the PDMS substrate and the glass negative into the plasma machine for cleaning. After cleaning, the PDMS substrate and the glass negative are bonded together to ensure that no air bubbles remain on the bonding surface, and then compacted with a heavy object.

Further, the preparation method of the microfluidic chip includes the following steps:

(1) Cleaning the single crystal silicon wafer: first rinse with deionized water, then rinse with absolute ethanol, then rinse with deionized water, dry with nitrogen after completion, and then place it on a microcomputer temperature-controlled heating table at 90 °C Set for 15 minutes;

(2) Gluing: apply positive photoresist on the surface of the single crystal silicon wafer, and use a spin coater of a glue spinner for spin coating, the rotation speed is 700r/min, and the spin coating time is 20s;

(3) Baking: Place the spin-coated monocrystalline silicon wafer on a microcomputer temperature-controlled heating table at 60°C for 15 minutes;

(4) Exposure: place the photolithography mask on the surface of the photoresist and expose it with an exposure machine;

(5) Development: Use developer to develop the exposed single crystal silicon wafer, then use deionized water to clean the surface of the single crystal silicon wafer, and dry it with nitrogen gas;

(6) Post-baking: place the single crystal silicon wafer on a microcomputer temperature-controlled heating table at 60°C for 40 minutes, so that the photoresist on the surface of the silicon wafer is completely cured, and a silicon-based mold is obtained;

(7) Constructing a microfluidic chip: First mix PDMS and curing agent in a volume ratio of 10:1 and stir evenly, pour it on a silicon-based mold, use a vacuum pump to remove air bubbles, and place it at 60°C for heating under microcomputer temperature control. Stand on the stage for one and a half hours to fully cure the PDMS, and then peel off the fully cured PDMS from the silicon-based mold to obtain the upper fluid channel layer of the microfluidic chip;

(8) Cleaning the glass negative: pass the glass negative in sequence, rinse with absolute ethanol and deionized water, and then dry it with nitrogen; the glass negative is the glass support layer of the microfluidic chip;

(9) Bonding: Put the PDMS substrate and the glass negative into the plasma machine for cleaning. After cleaning, the PDMS substrate and the glass negative are bonded within 1 minute and ensure that no air bubbles remain on the bonding surface, and then use a heavy object. After compaction, the bonding time was 12h, and the microfluidic chip was fabricated.

A microfluidic chip for high-throughput screening of nanoparticle transvascular transport, comprising a fluid channel layer and a fluid permeation layer connected with the fluid channel layer, wherein the lower end surface of the fluid channel layer is in contact with the upper end surface of the fluid permeation layer, The fluid channel layer is provided with a fluid channel, the fluid channel is arranged on the lower end face of the fluid channel layer, one end of the fluid channel is provided with a sample inlet, and the other end of the fluid channel is provided with a sample outlet; The fluid permeation layer is provided with a seepage outlet, and the middle part of the fluid channel is communicated with the seepage outlet through a gap channel. Using this technical solution, the drug-loaded nanoparticle solution is injected into the fluid channel from the injection port through the pressure applying device. During the circulation of the drug-loaded nanoparticle solution in the fluid channel, a part of the drug-loaded nanoparticle solution flows out from the sample outlet, and the other part of the drug-loaded nanoparticle solution flows out from the sample outlet. The drug nanoparticle solution will enter the exudate through the interstitial channel, and the drug-loaded nanoparticle solution in the exudate is collected. The above process is repeated by replacing the drug-loaded nanoparticle solution, and the drug-loaded nanoparticles that are more suitable for penetrating the tumor blood vessel wall can be screened by comparing the content of the drug-loaded nanoparticles in the collected solution.

The microfluidic chip is prepared by a molding method, including the following steps:

(1) Cleaning the single crystal silicon wafer: rinse with deionized water, blow dry with nitrogen after rinsing, and then place it on a heating table to stand;

(2) Glue coating: apply positive photoresist to the surface of the single crystal silicon wafer and spin it;

(3) Baking: put the spin-coated monocrystalline silicon wafer on the heating table and let it stand;

(4) Exposure: place the photolithography mask on the surface of the photoresist for exposure;

(5) Development: develop the exposed single crystal silicon wafer, then clean the surface of the single crystal silicon wafer with deionized water, and dry it with nitrogen;

(6) Post-baking: the single crystal silicon wafer is placed on a heating table to stand, so that the photoresist on the surface of the silicon wafer is completely cured, and a silicon-based mold is obtained;

(7) Constructing a microfluidic chip: First mix PDMS and curing agent in a volume ratio of 10:1 and stir evenly, pour it on the silicon-based mold of the fluid channel layer, remove air bubbles, and place it on a heating table. The PDMS is completely cured, and the fully cured PDMS is removed from the silicon-based mold of the fluid channel layer to obtain the fluid channel layer; then PDMS and the curing agent are mixed in a volume ratio of 15:1 and stirred evenly, Pour it on the silicon-based mold of the fluid-permeable layer, remove air bubbles, and place it on a heating table. When the PDMS is not fully cured, remove it from the silicon-based mold of the fluid-permeable layer, and then quickly align it with the fluid channel layer. close clamping;

(8) Thermal bonding: The fluid channel layer and fluid permeation layer that are clamped together are placed on a heating table and left to stand.

Further, the preparation method of the microfluidic chip includes the following steps:

(1) First rinse with deionized water, then rinse with absolute ethanol, then rinse with deionized water, dry with nitrogen after completion, and then place it on a microcomputer temperature-controlled heating table at 90 °C for 15 minutes;

(2) Gluing: apply positive photoresist on the surface of the single crystal silicon wafer, and use a spin coater of a glue spinner for spin coating, the rotation speed is 700r/min, and the spin coating time is 20s;

(3) Baking: Place the spin-coated monocrystalline silicon wafer on a microcomputer temperature-controlled heating table at 60°C for 15 minutes;

(4) Exposure: place the photolithography mask on the surface of the photoresist and expose it with an exposure machine;

(5) Development: Use developer to develop the exposed single crystal silicon wafer, then use deionized water to clean the surface of the single crystal silicon wafer, and dry it with nitrogen gas;

(6) Post-baking: place the single crystal silicon wafer on a microcomputer temperature-controlled heating table at 60°C for 40 minutes, so that the photoresist on the surface of the silicon wafer is completely cured, and a silicon-based mold is obtained;

(7) Constructing a microfluidic chip: First mix PDMS and curing agent in a volume ratio of 10:1 and stir evenly, pour it on the silicon-based mold of the fluid channel layer, remove air bubbles with a vacuum pump, and place it at 60°C The microcomputer temperature-controlled heating table was left standing for one and a half hours to fully cure the PDMS, and then the fully cured PDMS was removed from the silicon-based mold of the fluid channel layer to obtain the upper layer of the microfluidic chip; then PDMS and curing agent were used. After mixing in a volume ratio of 15:1 and stirring evenly, pour it onto the silicon-based mold of the fluid permeable layer, remove air bubbles with a vacuum pump, and place it on a microcomputer-controlled heating table at 60°C for 30 minutes. When cured, it was peeled from the silicon-based mold of the fluid permeable layer and then quickly clamped against the fluid channel layer.

(8) Thermal bonding: The fluid channel layer and fluid permeation layer that are clamped together are placed on a microcomputer temperature-controlled heating table at 80°C for 10 hours, and the fabrication of the microfluidic chip is completed.

Compared with the prior art, the present invention has the following beneficial effects: the microfluidic chip can be used to screen drug-loaded nanoparticles suitable for penetrating the blood vessel wall, and the screening accuracy is high, which can meet the requirements of modern drug-loaded nanoparticles to penetrate To meet the needs of vascular barrier medical research, the consumables in the entire screening process are only drug-loaded nanoparticle solution, so the cost of the drug screening process is low, and the preparation process of the microfluidic chip is relatively simple and low in production cost.

It can be seen that, compared with the prior art, the present invention has outstanding substantive features and significant progress, and the beneficial effects of its implementation are also obvious.

Description of drawings

1 is a schematic plan view of a microfluidic chip for high-throughput screening of nanoparticle transvascular transport in Example 1;

2 is a schematic three-dimensional structure diagram of a microfluidic chip for high-throughput screening of nanoparticle transvascular transport in Example 1;

Fig. 3 is the enlarged view of A part in Fig. 2;

FIG. 4 is a schematic structural diagram of a microfluidic chip for high-throughput screening of nanoparticle transvascular transport in Example 2. FIG.

In the figure: 1-injection port, 2-fluid channel I, 3-sample outlet, 4-exudation port, 5-gap channel, 6-fluid channel II, 7-fluid channel layer, 8-glass support layer, 9 - fluid permeable layer, 10 - fluid channel.

Detailed ways

In order to clearly illustrate the technical features of the present solution, the present solution will be described below through specific embodiments and in conjunction with the accompanying drawings.

Example 1

As shown in Figures 1-3, a microfluidic chip for high-throughput screening of nanoparticle transvascular transport is characterized by comprising a fluid channel layer 7 and a glass support layer 8 connected to the fluid channel layer 7. The said The lower end surface of the fluid channel layer 7 is in contact with the upper end surface of the glass support layer 8, the fluid channel layer 7 is provided with a fluid channel I2 and a fluid channel II6, and one end of the fluid channel I2 is provided with a sample inlet 1, and the The other end of the fluid channel I2 is provided with a sample outlet 3, the middle of the fluid channel I2 is communicated with the fluid channel II6 through the gap channel 5, and one end of the fluid channel II6 is provided with a seepage port 4. The drug-loaded nanoparticle solution is injected into the fluid channel I2 from the injection port 1 through the pressure applying device. When passing through the gap channel 5, a part of the drug-loaded nanoparticle solution will flow out from the sample outlet 3 through the fluid channel I2, and another part of the drug-loaded nanoparticle solution will flow out from the sample outlet 3. The solution enters the fluid channel II6 through the gap channel 5 and flows out from the permeation port 4, and the sample solution flowing out of the permeation port 4 is collected. The above process is repeated by replacing the drug-loaded nanoparticle solution, and the drug-loaded nanoparticles that are more suitable for penetrating the tumor blood vessel wall can be screened by comparing the content of the drug-loaded nanoparticles in the collected sample solution. The size of the interstitial channel 5 is adapted to the size of the hole in the vessel wall caused by the lack of basement membrane and adhesion proteins in the tumor vessel wall. Both the sample outlet 3 and the seepage outlet 4 can be led out through a conduit.

The microfluidic chip is prepared by a molding method, and the specific method is:

(1) Cleaning the single crystal silicon wafer: first rinse with deionized water, then rinse with absolute ethanol, then rinse with deionized water, dry with nitrogen after completion, and then place it on a microcomputer temperature-controlled heating table at 90 °C Set for 15 minutes;

(2) Gluing: apply positive photoresist on the surface of the single crystal silicon wafer, and use a spin coater of a glue spinner for spin coating, the rotation speed is 700r/min, and the spin coating time is 20s;

(3) Baking: Place the spin-coated monocrystalline silicon wafer on a microcomputer temperature-controlled heating table at 60°C for 15 minutes;

(4) Exposure: place the photolithography mask on the surface of the photoresist and expose it with an exposure machine;

(5) Development: Use developer to develop the exposed single crystal silicon wafer, then use deionized water to clean the surface of the single crystal silicon wafer, and dry it with nitrogen gas;

(6) Post-baking: place the single crystal silicon wafer on a microcomputer temperature-controlled heating table at 60°C for 40 minutes, so that the photoresist on the surface of the silicon wafer is completely cured, and a silicon-based mold is obtained;

(7) Constructing a microfluidic chip: First mix PDMS and curing agent in a volume ratio of 10:1 and stir evenly, pour it on a silicon-based mold, use a vacuum pump to remove air bubbles, and place it at 60°C for heating under microcomputer temperature control. Stand on the stage for one and a half hours to fully cure the PDMS, and then peel off the fully cured PDMS from the silicon-based mold to obtain the upper fluid channel layer of the microfluidic chip;

(8) Cleaning the glass negative: pass the glass negative in sequence, rinse with absolute ethanol and deionized water, and then dry it with nitrogen; the glass negative is the glass support layer of the microfluidic chip;

(9) Bonding: Put the PDMS substrate and the glass negative into the plasma machine for cleaning. After cleaning, the PDMS substrate and the glass negative are bonded within 1 minute and ensure that no air bubbles remain on the bonding surface, and then use a heavy object. After compaction, the bonding time was 12h, and the microfluidic chip was fabricated.

The PDMS is a polydimethylsilazane polymer, and the PDMS has good biocompatibility and optical properties. The PDMS and curing agent are Dow Corning SYLGARD184 silicone rubber.

Example 2

A microfluidic chip for high-throughput screening of nanoparticle transvascular transport, comprising a fluid channel layer 7 and a fluid permeation layer 9 connected to the fluid channel layer 7, the lower end face of the fluid channel layer 7 and the fluid permeation layer 9 The fluid channel layer 7 is provided with a fluid channel 10, the fluid channel 10 is arranged on the lower end surface of the fluid channel layer 7, and one end of the fluid channel 10 is provided with a sample inlet 1, and the fluid The other end of the channel 10 is provided with a sample outlet 3 ; the fluid permeable layer 9 is provided with a seepage outlet 4 , and the middle of the fluid channel 10 communicates with the seepage outlet 4 through a gap channel 5 . The drug-loaded nanoparticle solution is injected into the fluid channel 10 from the injection port 1 through the pressure applying device. During the flow of the drug-loaded nanoparticle solution in the fluid channel 10, a part of the drug-loaded nanoparticle solution flows out from the sample outlet 3, and the other part is drug-loaded. The nanoparticle solution will enter the infiltration port 4 through the interstitial channel 5, and the drug-loaded nanoparticle solution in the infiltration port will be collected. The above process is repeated by replacing the drug-loaded nanoparticle solution, and the drug-loaded nanoparticles that are more suitable for penetrating the tumor blood vessel wall can be screened by comparing the content of the drug-loaded nanoparticles in the collected solution. The size of the interstitial channel 5 can be adapted to the size of the hole in the vessel wall caused by the lack of basement membrane and adhesion proteins in the tumor vessel wall. Both the sample outlet 3 and the seepage outlet 4 can be led out through a conduit.

The microfluidic chip is prepared by a molding method, and the specific method is:

(1) First rinse with deionized water, then rinse with absolute ethanol, then rinse with deionized water, dry with nitrogen after completion, and then place it on a microcomputer temperature-controlled heating table at 90 °C for 15 minutes;

(2) Gluing: apply positive photoresist on the surface of the single crystal silicon wafer, and use a spin coater of a glue spinner for spin coating, the rotation speed is 700r/min, and the spin coating time is 20s;

(3) Baking: Place the spin-coated monocrystalline silicon wafer on a microcomputer temperature-controlled heating table at 60°C for 15 minutes;

(4) Exposure: place the photolithography mask on the surface of the photoresist and expose it with an exposure machine;

(5) Development: Use developer to develop the exposed single crystal silicon wafer, then use deionized water to clean the surface of the single crystal silicon wafer, and dry it with nitrogen gas;

(6) Post-baking: place the single crystal silicon wafer on a microcomputer temperature-controlled heating table at 60°C for 40 minutes, so that the photoresist on the surface of the silicon wafer is completely cured, and a silicon-based mold is obtained;

(7) Constructing a microfluidic chip: First mix PDMS and curing agent in a volume ratio of 10:1 and stir evenly, pour it on the silicon-based mold of the fluid channel layer, remove air bubbles with a vacuum pump, and place it at 60°C The microcomputer temperature-controlled heating table was left standing for one and a half hours to fully cure the PDMS, and then the fully cured PDMS was removed from the silicon-based mold of the fluid channel layer to obtain the upper layer of the microfluidic chip; then PDMS and curing agent were used. After mixing in a volume ratio of 15:1 and stirring evenly, pour it onto the silicon-based mold of the fluid permeable layer, remove air bubbles with a vacuum pump, and place it on a microcomputer-controlled heating table at 60°C for 30 minutes. When cured, it was peeled from the silicon-based mold of the fluid permeable layer and then quickly clamped against the fluid channel layer.

(8) Thermal bonding: The fluid channel layer and fluid permeation layer that are clamped together are placed on a microcomputer temperature-controlled heating table at 80°C for 10 hours, and the fabrication of the microfluidic chip is completed.

The PDMS is a polydimethylsilazane polymer, and the PDMS has good biocompatibility and optical properties. The PDMS and curing agent are Dow Corning SYLGARD184 silicone rubber.

The technical features that are not described in the present invention can be realized by or using the existing technology, and will not be repeated here. Of course, the above description is not a limitation of the present invention, and the present invention is not limited to the above-mentioned embodiments. Changes, modifications, additions or substitutions made by persons within the essential scope of the present invention shall also belong to the protection scope of the present invention.

Claims (8)

1. a kind of microfluidic chip of nanoparticle transvascular transport high-throughput screening, it is characterized in that: comprise fluid channel layer and the glass support layer that is connected with fluid channel layer, the lower end face of described fluid channel layer and glass support layer The upper end surface of the fluid channel layer is in contact with the upper end surface of the fluid channel layer. The fluid channel layer is provided with a fluid channel I and a fluid channel II. One end of the fluid channel I is provided with a sample inlet, and the other end of the fluid channel I is provided with a sample outlet. The middle part of the fluid channel I is communicated with the fluid channel II through a gap channel, and one end of the fluid channel II is provided with a seepage outlet. 2 . A method for preparing a microfluidic chip for high-throughput screening of nanoparticle transvascular transport as claimed in claim 1 , wherein the microfluidic chip is prepared by a molding method. 3 . 3. the preparation method of the microfluidic chip of nanoparticle transvascular transport high-throughput screening according to claim 2, is characterized in that: comprises the following steps: (1) Cleaning the single crystal silicon wafer: rinse with deionized water, blow dry with nitrogen after completion, and then place it on a heating table to stand; (2) Glue coating: apply positive photoresist to the surface of the single crystal silicon wafer and spin it; (3) Baking: put the spin-coated monocrystalline silicon wafer on the heating table and let it stand; (4) Exposure: place the photolithography mask on the surface of the photoresist for exposure; (5) Development: develop the exposed single crystal silicon wafer, then clean the surface of the single crystal silicon wafer with deionized water, and dry it with nitrogen; (6) Post-baking: the single crystal silicon wafer is placed on a heating table to stand, so that the photoresist on the surface of the silicon wafer is completely cured, and a silicon-based mold is obtained; (7) Constructing a microfluidic chip: First mix PDMS and curing agent in a volume ratio of 10:1 and stir evenly, pour it on a silicon-based mold, remove air bubbles, and place it on a heating table to allow PDMS to completely cure. , and then peel off the fully cured PDMS from the silicon-based mold to obtain the PDMS matrix, which is the fluid channel layer of the microfluidic chip; (8) Cleaning the glass negative film: Rinse the glass negative film with absolute ethanol and deionized water in sequence, and then dry it with nitrogen; (9) Bonding: Put the PDMS substrate and the glass negative into the plasma machine for cleaning. After cleaning, the PDMS substrate and the glass negative are bonded together to ensure that no air bubbles remain on the bonding surface, and then compacted with a heavy object. 4. the preparation method of the microfluidic chip of nanoparticle transvascular transport high-throughput screening according to claim 3, is characterized in that: comprises the following steps: (1) Cleaning the single crystal silicon wafer: first rinse with deionized water, then rinse with absolute ethanol, then rinse with deionized water, dry with nitrogen after completion, and then place it on a microcomputer temperature-controlled heating table at 90 °C Set for 15 minutes; (2) Gluing: apply positive photoresist on the surface of the single crystal silicon wafer, and use a spin coater of a glue spinner for spin coating, the rotation speed is 700r/min, and the spin coating time is 20s; (3) Baking: Place the spin-coated monocrystalline silicon wafer on a microcomputer temperature-controlled heating table at 60°C for 15 minutes; (4) Exposure: place the photolithography mask on the surface of the photoresist and expose it with an exposure machine; (5) Development: Use developer to develop the exposed single crystal silicon wafer, then use deionized water to clean the surface of the single crystal silicon wafer, and dry it with nitrogen gas; (6) Post-baking: place the single crystal silicon wafer on a microcomputer temperature-controlled heating table at 60°C for 40 minutes, so that the photoresist on the surface of the silicon wafer is completely cured, and a silicon-based mold is obtained; (7) Constructing a microfluidic chip: First mix PDMS and curing agent in a volume ratio of 10:1 and stir evenly, pour it on a silicon-based mold, use a vacuum pump to remove air bubbles, and place it at 60°C for heating under microcomputer temperature control. Stand on the stage for one and a half hours to fully cure the PDMS, and then peel off the fully cured PDMS from the silicon-based mold to obtain the upper fluid channel layer of the microfluidic chip; (8) Cleaning the glass negative: pass the glass negative in sequence, rinse with absolute ethanol and deionized water, and then dry it with nitrogen; (9) Bonding: Put the PDMS substrate and the glass negative into the plasma machine for cleaning. After cleaning, attach the PDMS substrate and the glass negative within 1 minute and ensure that no air bubbles remain on the bonding surface, and then use a heavy object. After compaction, the bonding time was 12h, and the microfluidic chip was fabricated. 5. a microfluidic chip for high-throughput screening of nanoparticle transvascular transport, is characterized in that: comprising a fluid channel layer and a fluid permeation layer connected with the fluid channel layer, the lower end face of the fluid channel layer and the fluid permeation layer The upper end surface of the layer is in contact, the fluid channel layer is provided with a fluid channel, the fluid channel is arranged on the lower end surface of the fluid channel layer, one end of the fluid channel is provided with a sample inlet, and the other end of the fluid channel is provided with There is a sample outlet; the fluid permeation layer is provided with a seepage outlet, and the middle part of the fluid channel is communicated with the seepage outlet through a gap channel. 6 . A method for preparing a microfluidic chip for high-throughput screening of nanoparticle transvascular transport as claimed in claim 5 , wherein the microfluidic chip is prepared by a molding method. 7 . 7. the preparation method of the microfluidic chip of nanoparticle transvascular transport high-throughput screening according to claim 6, is characterized in that: comprises the following steps: (1) Cleaning the single crystal silicon wafer: rinse with deionized water, blow dry with nitrogen after rinsing, and then place it on a heating table to stand; (2) Glue coating: apply positive photoresist to the surface of the single crystal silicon wafer and spin it; (3) Baking: put the spin-coated monocrystalline silicon wafer on the heating table and let it stand; (4) Exposure: place the photolithography mask on the surface of the photoresist for exposure; (5) Development: develop the exposed single crystal silicon wafer, then clean the surface of the single crystal silicon wafer with deionized water, and dry it with nitrogen; (6) Post-baking: the single crystal silicon wafer is placed on a heating table to stand, so that the photoresist on the surface of the silicon wafer is completely cured, and a silicon-based mold is obtained; (7) Constructing a microfluidic chip: First mix PDMS and curing agent in a volume ratio of 10:1 and stir evenly, pour it on the silicon-based mold of the fluid channel layer, remove air bubbles, and place it on a heating table. The PDMS is completely cured, and the fully cured PDMS is removed from the silicon-based mold of the fluid channel layer to obtain the fluid channel layer; then PDMS and the curing agent are mixed in a volume ratio of 15:1 and stirred evenly, Pour it on the silicon-based mold of the fluid-permeable layer, remove air bubbles, and place it on a heating table. When the PDMS is not fully cured, remove it from the silicon-based mold of the fluid-permeable layer, and then quickly align it with the fluid channel layer. close clamping; (8) Thermal bonding: The fluid channel layer and fluid permeation layer that are clamped together are placed on a heating table and left to stand. 8. the preparation method of the microfluidic chip of nanoparticle transvascular transport high-throughput screening according to claim 7, is characterized in that: comprises the following steps: (1) First rinse with deionized water, then rinse with absolute ethanol, then rinse with deionized water, dry with nitrogen after completion, and then place it on a microcomputer temperature-controlled heating table at 90 °C for 15 minutes; (2) Gluing: apply positive photoresist on the surface of the single crystal silicon wafer, and use a spin coater of a glue spinner for spin coating, the rotation speed is 700r/min, and the spin coating time is 20s; (3) Baking: Place the spin-coated monocrystalline silicon wafer on a microcomputer temperature-controlled heating table at 60°C for 15 minutes; (4) Exposure: place the photolithography mask on the surface of the photoresist and expose it with an exposure machine; (5) Development: Use developer to develop the exposed single crystal silicon wafer, then use deionized water to clean the surface of the single crystal silicon wafer, and dry it with nitrogen gas; (6) Post-baking: place the single crystal silicon wafer on a microcomputer temperature-controlled heating table at 60°C for 40 minutes, so that the photoresist on the surface of the silicon wafer is completely cured, and a silicon-based mold is obtained; (7) Constructing a microfluidic chip: First mix PDMS and curing agent in a volume ratio of 10:1 and stir evenly, pour it on the silicon-based mold of the fluid channel layer, remove air bubbles with a vacuum pump, and place it at 60°C The microcomputer temperature-controlled heating table was left standing for one and a half hours to fully cure the PDMS, and then the fully cured PDMS was removed from the silicon-based mold of the fluid channel layer to obtain the upper layer of the microfluidic chip; then PDMS and curing agent were used. After mixing in a volume ratio of 15:1 and stirring evenly, pour it onto the silicon-based mold of the fluid permeable layer, remove air bubbles with a vacuum pump, and place it on a microcomputer-controlled heating table at 60°C for 30 minutes. When curing, peel it off the silicon-based mold of the fluid permeable layer, and then quickly clamp it with the fluid channel layer; (8) Thermal bonding: The fluid channel layer and fluid permeation layer that are clamped together are placed on a microcomputer temperature-controlled heating table at 80°C for 10 hours, and the fabrication of the microfluidic chip is completed.

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