CN110773244A - Micro-fluidic chip for high-throughput screening of nano-particles in cross-vascular transport and preparation method thereof - Google Patents
Micro-fluidic chip for high-throughput screening of nano-particles in cross-vascular transport and preparation method thereof Download PDFInfo
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- B01L3/502761—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 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules
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- 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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Abstract
A high-throughput screening microfluidic chip for transporting nanoparticles across blood vessels and a preparation method thereof are provided, the microfluidic chip comprises a fluid channel layer and a glass supporting layer connected with the fluid channel layer, the lower end face of the fluid channel layer is in contact with the upper end face of the glass supporting layer, a fluid channel I and a fluid channel II are arranged in the fluid channel layer, one end of the fluid channel I is provided with a sample inlet, the other end of the fluid channel I is provided with a sample outlet, the middle 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 microfluidic chip can be used for screening drug-loaded nanoparticles suitable for penetrating through the vascular wall, and the screening accuracy is high.
Description
Technical Field
The invention relates to the technical field of applying a micro-fluidic chip technology to biomedical detection and screening, in particular to a micro-fluidic chip for high-throughput screening of nano-particle cross-vascular transport and a preparation method thereof.
Background
Tumors become the first killers threatening human health at present, and high-efficiency antitumor drugs become important requirements of healthy life of people. With the development of nanotechnology in recent years, nanoparticle drug-loaded therapy for tumors has been widely studied. By adopting the nano technology, the water solubility of the drug can be improved, the pharmacokinetic parameters of the drug in vivo can be improved, and the release speed of the drug can be controlled. However, the therapeutic effect is still not satisfactory as far as now. The reason for this is that the tumor microenvironment is different from the normal human environment in physicochemical properties, and is characterized by low oxygen, low pH, high osmotic pressure and high interstitial fluid pressure. It is also because of these characteristics that the drug cannot reach the tumor tissue at a uniform and effective concentration, resulting in insufficient local drug concentration, and this heterogeneous drug distribution is the main reason for the poor therapeutic effect of the drug-loaded nanoparticles. The key to the treatment of tumors is to study how to maximize the nanoparticles' reach to the tumors, so it is necessary to fully understand the nanoparticle transport process in the human body.
The transport of nanoparticles in the human body can be roughly divided into four stages, transport in blood vessels, penetration of the vessel wall into the interstitial space, transport in the interstitial space and entry into tumor cells. Wherein the transport through the vessel wall plays an important role in the whole transport process, and because the tumor vessel wall lacks basement membrane and adhesion protein, so that a plurality of holes appear on the vessel wall, the nano particles can penetrate through the holes. However, researchers have conducted few studies in this regard, and there is still a lack of clear understanding about the physicochemical properties of nanoparticles and the effect of the environment in blood vessels on their efficiency of penetrating the vessel wall. Therefore, the establishment of a device to screen for nanoparticles suitable for penetrating the vessel wall and a suitable vascular environment is a primary task in current tumor therapy research.
Microfluidic technology is an emerging interdisciplinary technology which is rapidly developed in recent years, and micro-machining and micro-operation which are difficult to be completed by a series of conventional methods can be realized by constructing micro-channels on a chip to process or operate micro-fluid. More accurate operation can be provided in the spatial dimension, and the dimension is closer to the actual dimension. Therefore, the microfluidic chip can be used as a platform for simulating the environment in a human body. At present, the research of screening the nano particles suitable for penetrating through the blood vessel wall and the suitable blood vessel environment by utilizing the microfluidic chip is still blank.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides the microfluidic chip for high-throughput screening of the cross-vascular transport of the nanoparticles and the preparation method thereof.
The scheme is realized by the following technical measures: a high-throughput screening microfluidic chip for transporting nanoparticles across blood vessels is characterized in that: including fluid passage layer and the glass supporting layer of being connected with the fluid passage layer, the lower terminal surface on fluid passage layer contacts with the up end of glass supporting layer, fluid passage I and fluid passage II are provided with in the fluid passage layer, the one end of fluid passage I is provided with the introduction port, the other end of fluid passage I is provided with the outlet, clearance passageway and II intercommunications of fluid passage are passed through to the middle part of fluid passage I, the one end of fluid passage II is provided with the oozing mouth. Medicine carrying nanoparticle solution is from the injection port injection fluid channel I through pressure application device, and when the process clearance passageway, partly medicine carrying nanoparticle solution can flow out from the play appearance mouth through fluid channel I, and another part medicine carrying nanoparticle solution passes through clearance passageway and gets into fluid channel II and flow out from the infiltration mouth, collects the sample solution of infiltration mouth outflow. The medicine carrying nano-particle solution is replaced, the above processes are repeated, the medicine carrying nano-particle content in the sample solution is collected in a contrast manner, and the medicine carrying nano-particles which are more suitable for penetrating through the wall of the tumor vessel can be screened.
The micro-fluidic chip is prepared by adopting a molding method and comprises the following steps:
(1) cleaning a monocrystalline silicon wafer: washing with deionized water, drying with nitrogen after the washing, and standing on a heating table;
(2) gluing: coating positive photoresist on the surface of a monocrystalline silicon wafer, and performing spin coating;
(3) baking: placing the spin-coated monocrystalline silicon wafer on a heating table for standing;
(4) exposure: placing the photoetching mask plate on the surface of the photoresist, and exposing;
(5) and (3) developing: developing the exposed monocrystalline silicon wafer, cleaning the surface of the monocrystalline silicon wafer by using deionized water, and drying by using nitrogen;
(6) post-baking: placing a monocrystalline silicon wafer on a heating table, and standing to completely cure the photoresist on the surface of the silicon wafer to obtain a silicon-based mold;
(7) constructing a micro-fluidic chip: firstly, mixing PDMS and a curing agent according to a volume ratio of 10:1, uniformly stirring, pouring the mixture on a silicon-based mold, removing bubbles, placing the mixture on a heating table, standing the mixture to completely cure the PDMS, and then removing the completely cured PDMS from the silicon-based mold to obtain a PDMS substrate, namely a fluid channel layer of the microfluidic chip;
(8) cleaning the glass negative plate: washing the glass negative film with absolute ethyl alcohol and deionized water in sequence, and then drying the glass negative film with nitrogen;
(9) bonding: putting the PDMS substrate and the glass substrate into a plasma machine for cleaning, after cleaning, attaching the PDMS substrate and the glass substrate and ensuring that no bubbles are left on the attaching surface, and then compacting by using a weight.
Further, the preparation method of the microfluidic chip comprises the following steps:
(1) cleaning a monocrystalline silicon wafer: washing with deionized water, washing with absolute ethyl alcohol, washing with deionized water, drying with nitrogen after washing, and standing on a microcomputer temperature-controlled heating table at 90 deg.C for 15 min;
(2) gluing: coating the positive photoresist on the surface of a monocrystalline silicon wafer, and spin-coating by using a spin coater of a spin coater at the rotation speed of 700r/min for 20 s;
(3) baking: placing the spin-coated monocrystalline silicon wafer on a microcomputer temperature control heating table at 60 ℃ and standing for 15 minutes;
(4) exposure: placing the photoetching mask plate on the surface of the photoresist, and exposing by using an exposure machine;
(5) and (3) developing: developing the exposed monocrystalline silicon wafer by using a developing solution, cleaning the surface of the monocrystalline silicon wafer by using deionized water, and drying by using nitrogen;
(6) post-baking: placing the monocrystalline silicon piece on a microcomputer temperature control heating table at 60 ℃ and standing for 40 minutes to completely cure the photoresist on the surface of the silicon piece to obtain a silicon-based mold;
(7) constructing a micro-fluidic chip: firstly, mixing PDMS and a curing agent according to a volume ratio of 10:1, uniformly stirring, pouring the mixture on a silicon-based mold, removing bubbles by using a vacuum pump, standing the mixture on a microcomputer temperature-controlled heating table at 60 ℃ for half an hour to completely cure the PDMS, and then removing the completely cured PDMS from the silicon-based mold to obtain an upper fluid channel layer of the microfluidic chip;
(8) cleaning the glass negative plate: sequentially washing the glass negative with absolute ethyl alcohol and deionized water, and then drying with nitrogen; the glass substrate is a glass supporting layer of the microfluidic chip;
(9) bonding: and putting the PDMS substrate and the glass substrate into a plasma machine for cleaning, after cleaning, adhering the PDMS substrate and the glass substrate within 1 minute and ensuring that no air bubbles are left on the adhering surface, then compacting by using a heavy object, and bonding for 12 hours to complete the manufacture of the microfluidic chip.
A micro-fluidic chip for high-throughput screening of nanoparticle cross-vascular transport comprises a fluid channel layer and a fluid permeation layer connected with the fluid channel layer, wherein the lower end face of the fluid channel layer is in contact with the upper end face of the fluid permeation layer, a fluid channel is arranged on the fluid channel layer, 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 permeable layer is provided with an seepage port, and the middle part of the fluid channel is communicated with the seepage port through a gap channel. Adopt this technical scheme, medicine carrying nanoparticle solution is put through pressure and is injected into fluid passage from the introduction port, and medicine carrying nanoparticle solution is at the in-process of fluid passage circulation, and partly medicine carrying nanoparticle solution flows out from the exit port, and another part medicine carrying nanoparticle solution can get into the infiltration mouth through clearance passageway, collects the medicine carrying nanoparticle solution in the infiltration mouth. The medicine carrying nano particle solution is replaced, the above processes are repeated, the medicine carrying nano particle content in the solution is collected in a contrast mode, and medicine carrying nano particles more suitable for penetrating through the wall of a tumor vessel can be screened out.
The micro-fluidic chip is prepared by adopting a molding method and comprises the following steps:
(1) cleaning a monocrystalline silicon wafer: washing with deionized water, drying with nitrogen after washing, and standing on a heating table;
(2) gluing: coating positive photoresist on the surface of a monocrystalline silicon wafer, and performing spin coating;
(3) baking: placing the spin-coated monocrystalline silicon wafer on a heating table for standing;
(4) exposure: placing the photoetching mask plate on the surface of the photoresist, and exposing;
(5) and (3) developing: developing the exposed monocrystalline silicon wafer, cleaning the surface of the monocrystalline silicon wafer by using deionized water, and drying by using nitrogen;
(6) post-baking: placing a monocrystalline silicon wafer on a heating table, and standing to completely cure the photoresist on the surface of the silicon wafer to obtain a silicon-based mold;
(7) constructing a micro-fluidic chip: firstly, mixing PDMS and a curing agent according to a volume ratio of 10:1, uniformly stirring, pouring the mixture on a silicon-based mold of a fluid channel layer, removing bubbles, placing the mixture on a heating table, standing the mixture to completely cure the PDMS, and then removing the completely cured PDMS from the silicon-based mold of the fluid channel layer to obtain the fluid channel layer; then mixing PDMS and a curing agent according to the volume ratio of 15:1, uniformly stirring, pouring the mixture on a silicon-based mold of a fluid permeation layer, removing air bubbles, placing the mixture on a heating table for standing, removing the PDMS from the silicon-based mold of the fluid permeation layer when the PDMS is not completely cured, and then quickly closing and clamping the PDMS and a fluid channel layer;
(8) thermal bonding: and placing the oppositely clamped fluid channel layer and the fluid permeation layer on a heating table for standing.
Further, the preparation method of the microfluidic chip comprises the following steps:
(1) washing with deionized water, washing with absolute ethyl alcohol, washing with deionized water, drying with nitrogen after washing, and standing on a microcomputer temperature-controlled heating table at 90 deg.C for 15 min;
(2) gluing: coating the positive photoresist on the surface of a monocrystalline silicon wafer, and spin-coating by using a spin coater of a spin coater at the rotation speed of 700r/min for 20 s;
(3) baking: placing the spin-coated monocrystalline silicon wafer on a microcomputer temperature control heating table at 60 ℃ and standing for 15 minutes;
(4) exposure: placing the photoetching mask plate on the surface of the photoresist, and exposing by using an exposure machine;
(5) and (3) developing: developing the exposed monocrystalline silicon wafer by using a developing solution, cleaning the surface of the monocrystalline silicon wafer by using deionized water, and drying by using nitrogen;
(6) post-baking: placing the monocrystalline silicon piece on a microcomputer temperature control heating table at 60 ℃ and standing for 40 minutes to completely cure the photoresist on the surface of the silicon piece to obtain a silicon-based mold;
(7) constructing a micro-fluidic chip: mixing PDMS and a curing agent according to a volume ratio of 10:1, uniformly stirring, pouring the mixture on a silicon-based mold of a fluid channel layer, removing bubbles by using a vacuum pumping pump, standing the mixture on a microcomputer temperature-control heating table at 60 ℃ for half an hour to completely cure the PDMS, and then removing the completely cured PDMS from the silicon-based mold of the fluid channel layer to obtain an upper layer of the microfluidic chip; then mixing PDMS and a curing agent according to a volume ratio of 15:1, uniformly stirring, pouring onto a silicon-based mold of the fluid permeation layer, removing air bubbles by using a vacuum pump, standing for 30 minutes on a microcomputer temperature control heating table at 60 ℃, removing the PDMS from the silicon-based mold of the fluid permeation layer when the PDMS is not completely cured, and then quickly closing and clamping with the fluid channel layer.
(8) Thermal bonding: and placing the oppositely clamped fluid channel layer and the fluid permeable layer on a microcomputer temperature control heating table at the temperature of 80 ℃ for standing for 10 hours, and finishing the manufacture of the microfluidic chip.
Compared with the prior art, the invention has the following beneficial effects: the microfluidic chip can be used for screening drug-loaded nanoparticles suitable for penetrating through a vascular wall, the screening accuracy is high, the requirement that modern drug-loaded nanoparticles penetrate through vascular barrier medical research can be met, only drug-loaded nanoparticle solution is consumed in the whole screening process, so the process cost of drug screening is low, the preparation process of the microfluidic chip is relatively simple, and the manufacturing cost is low.
Therefore, compared with the prior art, the invention has prominent substantive features and remarkable progress, and the beneficial effects of the implementation are also obvious.
Drawings
FIG. 1 is a schematic diagram of a planar structure of a microfluidic chip for high throughput screening of nanoparticles transported across blood vessels in example 1;
FIG. 2 is a schematic perspective view of a microfluidic chip for high throughput screening of nanoparticles transported across blood vessels in example 1;
FIG. 3 is an enlarged view of portion A of FIG. 2;
fig. 4 is a schematic structural diagram of the microfluidic chip for high-throughput screening of nanoparticles transported across blood vessels in example 2.
In the figure: 1-a sample inlet, 2-a fluid channel I, 3-a sample outlet, 4-a seepage outlet, 5-a gap channel, 6-a fluid channel II, 7-a fluid channel layer, 8-a glass supporting layer, 9-a fluid permeation layer and 10-a fluid channel.
Detailed Description
In order to clearly illustrate the technical features of the present solution, the following explains the present solution by way of specific embodiments and with reference to the accompanying drawings.
Example 1
As shown in fig. 1-3, a microfluidic chip for high-throughput screening of nanoparticle cross-vessel transport is characterized in that: including fluid channel layer 7 and the glass supporting layer 8 of being connected with fluid channel layer 7, the lower terminal surface of fluid channel layer 7 contacts with the up end of glass supporting layer 8, be provided with fluid channel I2 and fluid channel II 6 in the fluid channel layer 7, the one end of fluid channel I2 is provided with introduction port 1, the other end of fluid channel I2 is provided with out appearance mouth 3, the middle part of fluid channel I2 passes through clearance passageway 5 and fluid channel II 6 intercommunication, the one end of fluid channel II 6 is provided with ooze mouth 4. Medicine carrying nanoparticle solution is from introduction port 1 injection fluid channel I2 through pressure application device, and when the process clearance passageway 5, some medicine carrying nanoparticle solution can flow out from outlet port 3 through fluid channel I2, and another part medicine carrying nanoparticle solution passes through clearance passageway 5 and gets into fluid channel II 6 and flow out from oozing port 4, collects the sample solution that oozes out 4 outflow. The medicine carrying nano-particle solution is replaced, the above processes are repeated, the medicine carrying nano-particle content in the sample solution is collected in a contrast manner, and the medicine carrying nano-particles which are more suitable for penetrating through the wall of the tumor vessel can be screened. The size of the gap channel 5 is adapted to the size of the hole in the tumor vessel wall caused by the absence of basement membrane and adhesion proteins. The sample outlet 3 and the seepage outlet 4 can be led out through a conduit.
The micro-fluidic chip is prepared by adopting a molding method, and the specific method comprises the following steps:
(1) cleaning a monocrystalline silicon wafer: washing with deionized water, washing with absolute ethyl alcohol, washing with deionized water, drying with nitrogen after washing, and standing on a microcomputer temperature-controlled heating table at 90 deg.C for 15 min;
(2) gluing: coating the positive photoresist on the surface of a monocrystalline silicon wafer, and spin-coating by using a spin coater of a spin coater at the rotation speed of 700r/min for 20 s;
(3) baking: placing the spin-coated monocrystalline silicon wafer on a microcomputer temperature control heating table at 60 ℃ and standing for 15 minutes;
(4) exposure: placing the photoetching mask plate on the surface of the photoresist, and exposing by using an exposure machine;
(5) and (3) developing: developing the exposed monocrystalline silicon wafer by using a developing solution, cleaning the surface of the monocrystalline silicon wafer by using deionized water, and drying by using nitrogen;
(6) post-baking: placing the monocrystalline silicon piece on a microcomputer temperature control heating table at 60 ℃ and standing for 40 minutes to completely cure the photoresist on the surface of the silicon piece to obtain a silicon-based mold;
(7) constructing a micro-fluidic chip: firstly, mixing PDMS and a curing agent according to a volume ratio of 10:1, uniformly stirring, pouring the mixture on a silicon-based mold, removing bubbles by using a vacuum pump, standing the mixture on a microcomputer temperature-controlled heating table at 60 ℃ for half an hour to completely cure the PDMS, and then removing the completely cured PDMS from the silicon-based mold to obtain an upper fluid channel layer of the microfluidic chip;
(8) cleaning the glass negative plate: sequentially washing the glass negative with absolute ethyl alcohol and deionized water, and then drying with nitrogen; the glass substrate is a glass supporting layer of the microfluidic chip;
(9) bonding: and putting the PDMS substrate and the glass substrate into a plasma machine for cleaning, after cleaning, adhering the PDMS substrate and the glass substrate within 1 minute and ensuring that no air bubbles are left on the adhering surface, then compacting by using a heavy object, and bonding for 12 hours to complete the manufacture of the microfluidic chip.
The PDMS is a polydimethylsiloxane polymer, and has good biocompatibility and optical characteristics. The PDMS and the curing agent adopt American Dow Corning SYLGARD184 silicon rubber.
Example 2
A micro-fluidic chip for high-throughput screening of nanoparticle cross-blood vessel transport comprises a fluid channel layer 7 and a fluid permeation layer 9 connected with the fluid channel layer 7, wherein the lower end face of the fluid channel layer 7 is in contact with the upper end face of the fluid permeation layer 9, a fluid channel 10 is arranged on the fluid channel layer 7, the fluid channel 10 is arranged on the lower end face of the fluid channel layer 7, one end of the fluid channel 10 is provided with a sample inlet 1, and the other end of the fluid channel 10 is provided with a sample outlet 3; the fluid permeable layer 9 is provided with a seepage port 4, and the middle part of the fluid channel 10 is communicated with the seepage port 4 through a gap channel 5. Medicine carrying nanoparticle solution is from introduction port 1 injection fluid channel 10 through pressure application device, and medicine carrying nanoparticle solution is at the in-process of fluid channel 10 circulation, and partly medicine carrying nanoparticle solution flows out from outlet 3, and another part medicine carrying nanoparticle solution can get into oozing port 4 through clearance passageway 5, collects the interior medicine carrying nanoparticle solution of oozing port. The medicine carrying nano particle solution is replaced, the above processes are repeated, the medicine carrying nano particle content in the solution is collected in a contrast mode, and medicine carrying nano particles more suitable for penetrating through the wall of a tumor vessel can be screened out. The size of the gap tunnel 5 can be adapted to the size of the hole in the wall of the tumour vessel caused by the absence of basement membrane and adhesion proteins. The sample outlet 3 and the seepage outlet 4 can be led out through a conduit.
The micro-fluidic chip is prepared by adopting a molding method, and the specific method comprises the following steps:
(1) washing with deionized water, washing with absolute ethyl alcohol, washing with deionized water, drying with nitrogen after washing, and standing on a microcomputer temperature-controlled heating table at 90 deg.C for 15 min;
(2) gluing: coating the positive photoresist on the surface of a monocrystalline silicon wafer, and spin-coating by using a spin coater of a spin coater at the rotation speed of 700r/min for 20 s;
(3) baking: placing the spin-coated monocrystalline silicon wafer on a microcomputer temperature control heating table at 60 ℃ and standing for 15 minutes;
(4) exposure: placing the photoetching mask plate on the surface of the photoresist, and exposing by using an exposure machine;
(5) and (3) developing: developing the exposed monocrystalline silicon wafer by using a developing solution, cleaning the surface of the monocrystalline silicon wafer by using deionized water, and drying by using nitrogen;
(6) post-baking: placing the monocrystalline silicon piece on a microcomputer temperature control heating table at 60 ℃ and standing for 40 minutes to completely cure the photoresist on the surface of the silicon piece to obtain a silicon-based mold;
(7) constructing a micro-fluidic chip: mixing PDMS and a curing agent according to a volume ratio of 10:1, uniformly stirring, pouring the mixture on a silicon-based mold of a fluid channel layer, removing bubbles by using a vacuum pumping pump, standing the mixture on a microcomputer temperature-control heating table at 60 ℃ for half an hour to completely cure the PDMS, and then removing the completely cured PDMS from the silicon-based mold of the fluid channel layer to obtain an upper layer of the microfluidic chip; then mixing PDMS and a curing agent according to a volume ratio of 15:1, uniformly stirring, pouring onto a silicon-based mold of the fluid permeation layer, removing air bubbles by using a vacuum pump, standing for 30 minutes on a microcomputer temperature control heating table at 60 ℃, removing the PDMS from the silicon-based mold of the fluid permeation layer when the PDMS is not completely cured, and then quickly closing and clamping with the fluid channel layer.
(8) Thermal bonding: and placing the oppositely clamped fluid channel layer and the fluid permeable layer on a microcomputer temperature control heating table at the temperature of 80 ℃ for standing for 10 hours, and finishing the manufacture of the microfluidic chip.
The PDMS is a polydimethylsiloxane polymer, and has good biocompatibility and optical characteristics. The PDMS and the curing agent adopt American Dow Corning SYLGARD184 silicon rubber.
The technical features of the present invention that are not described in the present invention can be implemented by or using the prior art, and are not described herein again, of course, the above description is not limited to the present invention, and the present invention is not limited to the above embodiments, and variations, modifications, additions or substitutions that are made by those skilled in the art within the spirit and scope of the present invention should also fall within the protection scope of the present invention.
Claims (8)
1. A high-throughput screening microfluidic chip for transporting nanoparticles across blood vessels is characterized in that: including fluid passage layer and the glass supporting layer of being connected with the fluid passage layer, the lower terminal surface on fluid passage layer contacts with the up end of glass supporting layer, fluid passage I and fluid passage II are provided with in the fluid passage layer, the one end of fluid passage I is provided with the introduction port, the other end of fluid passage I is provided with the outlet, clearance passageway and II intercommunications of fluid passage are passed through to the middle part of fluid passage I, the one end of fluid passage II is provided with the oozing mouth.
2. A method for preparing a microfluidic chip for high throughput screening of nanoparticles transported across blood vessels as claimed in claim 1, wherein the method comprises the following steps: the microfluidic chip is prepared by a molding method.
3. The method for preparing the microfluidic chip for high-throughput screening of nanoparticles transported across blood vessels according to claim 2, which is characterized in that: the method comprises the following steps:
(1) cleaning a monocrystalline silicon wafer: washing with deionized water, drying with nitrogen after the washing, and standing on a heating table;
(2) gluing: coating positive photoresist on the surface of a monocrystalline silicon wafer, and performing spin coating;
(3) baking: placing the spin-coated monocrystalline silicon wafer on a heating table for standing;
(4) exposure: placing the photoetching mask plate on the surface of the photoresist, and exposing;
(5) and (3) developing: developing the exposed monocrystalline silicon wafer, cleaning the surface of the monocrystalline silicon wafer by using deionized water, and drying by using nitrogen;
(6) post-baking: placing a monocrystalline silicon wafer on a heating table, and standing to completely cure the photoresist on the surface of the silicon wafer to obtain a silicon-based mold;
(7) constructing a micro-fluidic chip: firstly, mixing PDMS and a curing agent according to a volume ratio of 10:1, uniformly stirring, pouring the mixture on a silicon-based mold, removing bubbles, placing the mixture on a heating table, standing the mixture to completely cure the PDMS, and then removing the completely cured PDMS from the silicon-based mold to obtain a PDMS substrate, namely a fluid channel layer of the microfluidic chip;
(8) cleaning the glass negative plate: washing the glass negative film with absolute ethyl alcohol and deionized water in sequence, and then drying the glass negative film with nitrogen;
(9) bonding: putting the PDMS substrate and the glass substrate into a plasma machine for cleaning, after cleaning, attaching the PDMS substrate and the glass substrate and ensuring that no bubbles are left on the attaching surface, and then compacting by using a weight.
4. The method for preparing the microfluidic chip for high-throughput screening of nanoparticles transported across blood vessels according to claim 3, wherein the method comprises the following steps: the method comprises the following steps:
(1) cleaning a monocrystalline silicon wafer: washing with deionized water, washing with absolute ethyl alcohol, washing with deionized water, drying with nitrogen after washing, and standing on a microcomputer temperature-controlled heating table at 90 deg.C for 15 min;
(2) gluing: coating the positive photoresist on the surface of a monocrystalline silicon wafer, and spin-coating by using a spin coater of a spin coater at the rotation speed of 700r/min for 20 s;
(3) baking: placing the spin-coated monocrystalline silicon wafer on a microcomputer temperature control heating table at 60 ℃ and standing for 15 minutes;
(4) exposure: placing the photoetching mask plate on the surface of the photoresist, and exposing by using an exposure machine;
(5) and (3) developing: developing the exposed monocrystalline silicon wafer by using a developing solution, cleaning the surface of the monocrystalline silicon wafer by using deionized water, and drying by using nitrogen;
(6) post-baking: placing the monocrystalline silicon piece on a microcomputer temperature control heating table at 60 ℃ and standing for 40 minutes to completely cure the photoresist on the surface of the silicon piece to obtain a silicon-based mold;
(7) constructing a micro-fluidic chip: firstly, mixing PDMS and a curing agent according to a volume ratio of 10:1, uniformly stirring, pouring the mixture on a silicon-based mold, removing bubbles by using a vacuum pump, standing the mixture on a microcomputer temperature-controlled heating table at 60 ℃ for half an hour to completely cure the PDMS, and then removing the completely cured PDMS from the silicon-based mold to obtain an upper fluid channel layer of the microfluidic chip;
(8) cleaning the glass negative plate: sequentially washing the glass negative with absolute ethyl alcohol and deionized water, and then drying with nitrogen;
(9) bonding: and putting the PDMS substrate and the glass substrate into a plasma machine for cleaning, after cleaning, adhering the PDMS substrate and the glass substrate within 1 minute and ensuring that no air bubbles are left on the adhering surface, then compacting by using a heavy object, and bonding for 12 hours to complete the manufacture of the microfluidic chip.
5. A high-throughput screening microfluidic chip for transporting nanoparticles across blood vessels is characterized in that: the device comprises a fluid channel layer and a fluid permeation layer connected with the fluid channel layer, wherein the lower end face of the fluid channel layer is contacted with the upper end face of the fluid permeation layer, a fluid channel is arranged on the fluid channel layer and 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 permeable layer is provided with an seepage port, and the middle part of the fluid channel is communicated with the seepage port through a gap channel.
6. A method for preparing the microfluidic chip for high-throughput screening of nanoparticle cross-vessel transport as claimed in claim 5, which is characterized in that: the microfluidic chip is prepared by a molding method.
7. The method for preparing the microfluidic chip for high-throughput screening of nanoparticle cross-vascular transport according to claim 6, wherein the method comprises the following steps: the method comprises the following steps:
(1) cleaning a monocrystalline silicon wafer: washing with deionized water, drying with nitrogen after washing, and standing on a heating table;
(2) gluing: coating positive photoresist on the surface of a monocrystalline silicon wafer, and performing spin coating;
(3) baking: placing the spin-coated monocrystalline silicon wafer on a heating table for standing;
(4) exposure: placing the photoetching mask plate on the surface of the photoresist, and exposing;
(5) and (3) developing: developing the exposed monocrystalline silicon wafer, cleaning the surface of the monocrystalline silicon wafer by using deionized water, and drying by using nitrogen;
(6) post-baking: placing a monocrystalline silicon wafer on a heating table, and standing to completely cure the photoresist on the surface of the silicon wafer to obtain a silicon-based mold;
(7) constructing a micro-fluidic chip: firstly, mixing PDMS and a curing agent according to a volume ratio of 10:1, uniformly stirring, pouring the mixture on a silicon-based mold of a fluid channel layer, removing bubbles, placing the mixture on a heating table, standing the mixture to completely cure the PDMS, and then removing the completely cured PDMS from the silicon-based mold of the fluid channel layer to obtain the fluid channel layer; then mixing PDMS and a curing agent according to the volume ratio of 15:1, uniformly stirring, pouring the mixture on a silicon-based mold of a fluid permeation layer, removing air bubbles, placing the mixture on a heating table for standing, removing the PDMS from the silicon-based mold of the fluid permeation layer when the PDMS is not completely cured, and then quickly closing and clamping the PDMS and a fluid channel layer;
(8) thermal bonding: and placing the oppositely clamped fluid channel layer and the fluid permeation layer on a heating table for standing.
8. The method for preparing the microfluidic chip for high throughput screening of nanoparticles transported across blood vessels according to claim 7, wherein the method comprises the following steps: the method comprises the following steps:
(1) washing with deionized water, washing with absolute ethyl alcohol, washing with deionized water, drying with nitrogen after washing, and standing on a microcomputer temperature-controlled heating table at 90 deg.C for 15 min;
(2) gluing: coating the positive photoresist on the surface of a monocrystalline silicon wafer, and spin-coating by using a spin coater of a spin coater at the rotation speed of 700r/min for 20 s;
(3) baking: placing the spin-coated monocrystalline silicon wafer on a microcomputer temperature control heating table at 60 ℃ and standing for 15 minutes;
(4) exposure: placing the photoetching mask plate on the surface of the photoresist, and exposing by using an exposure machine;
(5) and (3) developing: developing the exposed monocrystalline silicon wafer by using a developing solution, cleaning the surface of the monocrystalline silicon wafer by using deionized water, and drying by using nitrogen;
(6) post-baking: placing the monocrystalline silicon piece on a microcomputer temperature control heating table at 60 ℃ and standing for 40 minutes to completely cure the photoresist on the surface of the silicon piece to obtain a silicon-based mold;
(7) constructing a micro-fluidic chip: mixing PDMS and a curing agent according to a volume ratio of 10:1, uniformly stirring, pouring the mixture on a silicon-based mold of a fluid channel layer, removing bubbles by using a vacuum pumping pump, standing the mixture on a microcomputer temperature-control heating table at 60 ℃ for half an hour to completely cure the PDMS, and then removing the completely cured PDMS from the silicon-based mold of the fluid channel layer to obtain an upper layer of the microfluidic chip; then mixing PDMS and a curing agent according to a volume ratio of 15:1, uniformly stirring, pouring the mixture on a silicon-based mold of a fluid permeation layer, removing air bubbles by using a vacuum pumping pump, standing the mixture on a microcomputer temperature control heating table at 60 ℃ for 30 minutes, removing the PDMS from the silicon-based mold of the fluid permeation layer when the PDMS is not completely cured, and then quickly closing and clamping the PDMS with a fluid channel layer;
(8) thermal bonding: and placing the oppositely clamped fluid channel layer and the fluid permeable layer on a microcomputer temperature control heating table at the temperature of 80 ℃ for standing for 10 hours, and finishing the manufacture of the microfluidic chip.
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