A 3D printed resin-based flip chip

Abstract

The present invention relates to the field of microfluidic chip technology, and in particular to a 3D printed resin-based flip chip. The flip chip of the present invention is prepared by a thermoplastic printing process to form a core area, a front fluid microchannel and a back fluid microchannel, a front flow-through liquid reservoir and a back lateral liquid reservoir, and a field of view window is formed by bonding with transparent 3D printed upper and lower layers, thereby completing the structural production; during the sampling process, the sample in the front flow-through liquid reservoir and the sample in the back lateral liquid reservoir are independent of each other. After the sampling is completed, the two samples are mixed by flipping the chip, and the reaction is combined. Through structural design, the sample volume in the detection pool can be automatically adjusted to perform different concentration gradient detections, and multi-target simultaneous detection is also achieved through parallel design. The preparation process of the present invention is simple, the batch-prepared chips have high consistency and repeatability, and the cost is low, the functionality is extremely strong, and it has good promotion and practicality.

Description

Technical Field

[0001] The present invention relates to the technical field of microfluidic chips, in particular to a 3D printed resin-based flip chip. Background Art

[0002] Microfluidic chip analysis systems are easy to integrate and portable, which provides them with broad application prospects in biomedicine, environmental detection and protection. With the development of various ultra-early cancer marker screening methods, fluorescent chips made by combining microfluidic methods with biological detection can effectively detect markers, but there are still problems in many aspects. Most of the current research focuses on a single tumor marker, and it is difficult for a single marker to meet the requirements of sensitivity and specificity at the same time. Therefore, it is of great significance to find a fluorescent chip that can detect multiple markers simultaneously and efficiently and accurately. At this stage, fluorescent chips still have the problems of high cost, low chip integration and sensitivity. How to reduce costs and improve the integration and detection sensitivity of fluorescent chips is also an urgent problem that needs to be solved. Summary of the Invention

[0003] The purpose of the present invention is to provide a 3D printed resin-based flip chip, which can realize the simultaneous detection of multiple targets and the different concentration gradient testing of a single target through the flip chip structure design.

[0004] To achieve this object, the present invention adopts the following technical solutions:

[0005] A first aspect of the present invention provides a 3D-printed resin-based flip chip, comprising an upper layer, a core region, and a lower layer bonded sequentially from top to bottom; the core region is provided with at least one group of reaction units, each group of reaction units comprising a front fluid microchannel and a rear fluid microchannel;

[0006] A plurality of flow-through liquid reservoirs are provided on the front fluid microchannel, wherein the front fluid microchannel horizontally penetrates the central region of the flow-through liquid reservoir, and the inlet and outlet of the front fluid microchannel are symmetrically distributed relative to the flow-through liquid reservoir; a plurality of lateral liquid reservoirs are provided on one side of the reverse fluid microchannel, wherein the reverse fluid microchannel does not penetrate the central region of the lateral liquid reservoir, and the inlet and outlet of the reverse fluid microchannel are located beside the lateral liquid reservoir;

[0007] The flow-through liquid reservoir and the lateral liquid reservoir are two cylinders integrally connected at the top and bottom and with interconnected inner cavities; the ratio of the width of the front fluid microchannel to the radius of the cylindrical cavity of the flow-through liquid reservoir is 1:2-4; the inner surface of the cylindrical cavity of the flow-through liquid reservoir is hydrophilic; the ratio of the width of the reverse fluid microchannel to the radius of the cylindrical cavity of the lateral liquid reservoir is 1:2-4; the inner surface of the cylindrical cavity of the lateral liquid reservoir is hydrophilic;

[0008] The upper layer is provided with a front sampling groove and a front sampling port communicating with the front fluid microchannel, and a rear sampling port communicating with the rear fluid microchannel; the lower layer is provided with a rear sampling groove and a rear sampling port communicating with the rear fluid microchannel, and a front sampling port communicating with the front fluid microchannel.

[0009] In some embodiments of the present invention, the inner diameter of the cylindrical cavity of the flow-through liquid reservoir is smaller than the inner diameter of the cylindrical cavity of the lateral liquid reservoir, and the inner cavity of the flow-through liquid reservoir is connected to the inner cavity of the lateral liquid reservoir through a fluid channel, and the fluid channel is a through hole provided at the center position of the top surface of the lateral liquid reservoir.

[0010] In some embodiments of the present invention, the front fluid microchannel and the reverse fluid microchannel in the core area are horizontally distributed; one end of the front fluid microchannel is provided with a vertically upward front sample injection channel, and the other end is provided with a vertically downward front sample outlet channel; one end of the reverse fluid microchannel is provided with a vertically downward reverse sample injection channel, and the other end is provided with a vertically upward reverse sample outlet channel; the front sample injection groove is located below the upper layer and is perpendicular to the horizontal direction of the front sample injection channel; the reverse sample injection groove is located below the lower layer and is perpendicular to the horizontal direction of the reverse sample injection channel.

[0011] In some embodiments of the present invention, the flow-through liquid reservoir and the lateral liquid reservoir are both distributed in a 1×N linear array, the cylindrical cavity radius and depth of the N flow-through liquid reservoirs are the same, the depth of the N lateral liquid reservoirs is the same, and the cylindrical cavity radius increases successively, so as to form a concentration gradient for the same target object by one injection.

[0012] In some embodiments of the present invention, the flow-through liquid reservoir and the lateral liquid reservoir are both distributed in a 1×N linear array, the cylindrical cavity radius and depth of the N flow-through liquid reservoirs are the same, and the cylindrical cavity radius of the N lateral liquid reservoirs is the same, and the depth increases successively, so as to form a concentration gradient for a single injection of the same target object.

[0013] In some embodiments of the present invention, the core area is provided with M groups of reaction units, and the flow-through liquid reservoirs and lateral liquid reservoirs of each group of reaction units are distributed in a 1×N linear array for simultaneous quantitative detection of multiple targets.

[0014] A second aspect of the present invention provides a method for preparing the 3D printed resin-based flip chip, comprising the following steps:

[0015] Step 1: Use 3D mapping software to perform regional modeling to obtain three-dimensional models of the upper layer, core area, and lower layer;

[0016] Step 2: Use a 3D printer to print a three-dimensional model of the upper layer, core area, and lower layer to obtain a printed structure;

[0017] Step 3: Cut off the base of the printed structure and use an organic solvent to remove surface impurities;

[0018] Step 4: UV curing the printed structure after impurities removal;

[0019] Step 5: Use adhesive tape to punch holes and assemble the printed structure.

[0020] The third aspect of the present invention provides the application of the 3D printed resin-based flip chip in tumor marker detection.

[0021] A fourth aspect of the present invention provides a method for using the 3D printed resin-based flip chip, comprising the following steps:

[0022] S1. Inject sample A from the front inlet of the flip chip. Sample A enters the front fluid microchannel and the flow-through reservoir from the front inlet under pressure. Excess sample A is discharged from the front outlet under gravity. Inject sample B from the back inlet of the chip. Sample B enters the back fluid microchannel and the lateral reservoir from the back inlet under pressure.

[0023] S2. Flip the chip 180°. During the flipping process, sample A and sample B are mixed through the fluid channel between the flow-through liquid reservoir and the lateral liquid reservoir to react.

[0024] Compared with the prior art, the present invention has the following beneficial effects:

[0025] 1. The 3D-printed resin-based flip chip provided by the present invention has high integration and small area. It can be mass-produced by 3D printing using transparent resin as the printing base material. The chip has high consistency and repeatability, which reduces the chip cost. In addition, the size, depth and gap size of the observation area can be adjusted and are not affected by the process.

[0026] 2. The 3D-printed resin-based flip chip provided by the present invention, through the structural design of the fluid microchannel and the liquid reservoir and the hydrophilic treatment of the inner surface of the liquid reservoir cavity, ensures that during the sampling process, the sample in the front flow-through liquid reservoir and the sample in the rear lateral liquid reservoir remain independent of each other. After the sampling is completed, the two samples are mixed and reacted by flipping the chip. The liquid reservoir array distribution design allows different concentration gradients to be obtained with a single injection. The parallel design of multiple reaction units enables the simultaneous detection of multiple analytes and their respective concentration gradients. Compared with the current well plate test kit, it is cheaper, simpler and faster, smaller in size, and more integrated.

[0027] 3. The present invention uses adhesive stickers to seal the sample to be tested in the cavity, isolating it from the outside world and helping to prevent sample contamination. It can produce low-cost, easy-to-operate, highly sensitive, and high-precision 3D printed chips for detecting a variety of targets. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 This is a process flow chart for preparing the flip chip of the present invention.

[0029] Figure 2 This is a schematic diagram of the structure of an equal-depth flip chip array with the same radius for the flow-through liquid reservoir and the lateral liquid reservoir of the present invention.

[0030] Figure 3 This is a schematic diagram of the structure of a non-equal-depth flip chip with an array of lateral liquid storage tanks of the same radius according to the present invention.

[0031] Figure 4 This is a schematic diagram of an array of equal-depth flip-type chip structures in which the flow-through liquid reservoirs have the same radius and the lateral liquid reservoirs have different radii.

[0032] Figure 5 Schematic diagram of the chip structure for simultaneous detection of multiple targets of the present invention.

[0033] Figure 6 It is a schematic diagram of the three-dimensional structure of the flip chip of the present invention.

[0034] Figure 7 It is a schematic diagram of the top view of the flip chip of the present invention.

[0035] Figure 8 This is a physical picture of the flip chip used for different concentration gradient detection in the present invention.

[0036] Figure 9 The flip chip of the present invention is used to detect cysteine.

[0037] Explanation of the accompanying symbols: 1-upper layer; 2-core area; 3-lower layer; 11-front sampling groove; 12-front sampling port; 13-rear sampling port; 21-front fluid microchannel; 22-rear fluid microchannel; 23-front sampling channel; 24-front sampling channel; 25-flow-through liquid storage tank; 26-rear sampling channel; 27-rear sampling channel; 28-lateral liquid storage tank; 29-fluid channel; 31-rear sampling groove; 32-rear sampling port; 33-front sampling port. DETAILED DESCRIPTION

[0038] The following describes in detail the 3D printed resin-based flip chip of the present invention, its preparation method, and application.

[0039] In the flow-through liquid storage tank of the present invention, the liquid flow channel horizontally passes through the central area of ​​the liquid storage tank, and the inlet and outlet of the flow channel are symmetrically distributed relative to the liquid storage tank.

[0040] In the lateral liquid storage tank of the present invention, the inlet and outlet of the fluid flow channel are located on the side of the liquid storage tank, and the flow channel does not pass through the central area of ​​the liquid storage tank.

[0041] The first aspect of the present invention provides a 3D printed resin-based flip chip, the structure of which is as follows: Figure 6 and Figure 7 As shown, the flip chip comprises an upper layer 1, a core region 2, and a lower layer 3 bonded sequentially from top to bottom. In some embodiments of the present invention, the bonding is achieved by punching holes in an adhesive tape to achieve integrated assembly of the upper layer 1, the core region 2, and the lower layer 3. The core region 2 is provided with at least one set of reaction units, each of which includes a front-side fluid microchannel 21 and a back-side fluid microchannel 22.

[0042] A plurality of flow-through liquid reservoirs 25 are provided on the front fluid microchannel 21 . The front fluid microchannel 21 horizontally passes through the central area of ​​the flow-through liquid reservoir 25 . The inlet and outlet of the front fluid microchannel 21 are symmetrically distributed relative to the flow-through liquid reservoir 25 .

[0043] A plurality of lateral liquid reservoirs 28 are provided on one side of the reverse fluid microchannel 22 . The reverse fluid microchannel 22 does not penetrate the central area of ​​the lateral liquid reservoir 28 . The inlet and outlet of the reverse fluid microchannel 22 are located beside the lateral liquid reservoir 28 .

[0044] The flow-through liquid reservoir 25 and the lateral liquid reservoir 28 are two cylinders connected in an integral manner from top to bottom and with their inner cavities communicating with each other; the ratio of the width of the front fluid microchannel 21 to the radius of the cylindrical cavity of the flow-through liquid reservoir 25 is 1:2-4; the inner surface of the cylindrical cavity of the flow-through liquid reservoir 25 is hydrophilic; the ratio of the width of the reverse fluid microchannel 22 to the radius of the cylindrical cavity of the lateral liquid reservoir 28 is 1:2-4; the inner surface of the cylindrical cavity of the lateral liquid reservoir 28 is hydrophilic. Through the above-mentioned structural design and surface treatment process, when the flip chip is placed face up, the sample solution in the flow-through liquid reservoir 25 can be adsorbed on the inner wall of the liquid reservoir and will not fall off, thereby avoiding contact with the sample solution in the lateral liquid reservoir 28 during the injection process. When the flip chip is flipped (oscillation can also be used), the sample solution in the inner cavity of the flow-through liquid reservoir 25 is mixed with the sample solution in the lateral liquid reservoir 28.

[0045] In the present invention, the flow-through liquid reservoir 25 and the corresponding lateral liquid reservoir 28 together form a detection area; when a fluorescent probe is used for detection, the lateral liquid reservoir 28 can be used as a fluorescence observation area.

[0046] The upper layer 1 is provided with a front sampling groove 11 communicating with the front fluid microchannel 21, a front sampling port 12 and a rear sampling port 13 communicating with the rear fluid microchannel 22; the lower layer 3 is provided with a rear sampling groove 31 communicating with the rear fluid microchannel 22, a rear sampling port 32 and a front sampling port 33 communicating with the front fluid microchannel 21.

[0047] 3D printing has good micromachining performance. The present invention uses 3D modeling software to design microfluidic channels and liquid reservoirs. Due to the flexibility of 3D printing, the number of reaction units, the number, distribution and depth of liquid reservoirs can be adjusted according to needs.

[0048] In the present invention, the hydrophilic treatment can be performed by surface plasma treatment to modify hydrophilic groups such as hydroxyl groups and amino groups on the surface of the fluid microchannel and the liquid reservoir, or by surfactant treatment, such as Triton solution, to improve the hydrophilicity of the surface of the fluid microchannel and the liquid reservoir.

[0049] In some embodiments of the present invention, the inner diameter of the cylindrical cavity of the flow-through liquid reservoir 25 is smaller than the inner diameter of the cylindrical cavity of the lateral liquid reservoir 28. The inner cavity of the flow-through liquid reservoir 25 and the inner cavity of the lateral liquid reservoir 28 are connected through a fluid channel 29. The fluid channel 29 is a through hole provided at the center of the top surface of the lateral liquid reservoir 28. Preferably, the fluid channel 29 is a circular hole provided at the center of the top surface of the lateral liquid reservoir 28. The radius of the circular hole is the same as the radius of the bottom surface of the cylindrical cavity of the flow-through liquid reservoir 25, that is, the flow-through liquid reservoir 25 is a bottomless cylinder ( Figure 6 As shown), the cylindrical cavity of the flow-through liquid reservoir 25 is directly connected to the cylindrical cavity of the lateral liquid reservoir 28.

[0050] In some embodiments of the present invention, Figure 6 and Figure 7 As shown, the front fluid microchannel 21 and the reverse fluid microchannel 22 in the core area are horizontally distributed; one end of the front fluid microchannel 21 is provided with a vertically upward front sample injection channel 23, and the other end is provided with a vertically downward front sample outlet channel 24; one end of the reverse fluid microchannel 22 is provided with a vertically downward reverse sample injection channel 26, and the other end is provided with a vertically upward reverse sample outlet channel 27; the front sample injection groove 11 is located below the upper layer 1, and is perpendicular to the horizontal direction of the front sample injection channel 23; the reverse sample injection groove 31 is located below the lower layer 3, and is perpendicular to the horizontal direction of the reverse sample injection channel 26.

[0051] The working principle of the flip chip of the present invention is as follows:

[0052] First, the chip is subjected to negative pressure treatment. When the front side is sampled, the probe sample is injected from the front sampling port 12 with a sampler. Due to the pressure difference, the probe sample will enter the front fluid microchannel 21 from the front sampling port 12 through the front sampling channel 23 under the action of the air pump or the pipette gun. Since the present invention controls the radius ratio of the front fluid microchannel 21 to the flow-through liquid reservoir 25 at 1:2-4, and performs hydrophilic treatment on the inner surface of the cavity of the flow-through liquid reservoir 25, the front fluid microchannel 21 is connected to the flow-through liquid reservoir 25. After the probe sample fills the first flow-through liquid reservoir 25, it enters the second flow-through liquid reservoir 25 through the front fluid microchannel 21. In this way, the remaining flow-through liquid reservoirs 25 are filled and maintained in the flow-through liquid reservoir 25 in turn. The excess probe sample flows out from the front sample outlet 33 through the front sample outlet channel 24 under the action of gravity ( Figure 6 The unidirectional arrow in the figure indicates the flow direction of the probe sample). During reverse injection, the sample to be tested is injected from the reverse injection port 32 using a sampler. Due to the pressure difference, the sample to be tested will enter the reverse fluid microchannel 22 from the reverse injection port 32 through the reverse injection channel 26 under the action of an air pump or a pipette. Since the present invention controls the radius ratio of the reverse fluid microchannel 22 to the lateral liquid reservoir 28 to be 1:2-4, and performs a hydrophilic treatment on the inner surface of the cavity of the lateral liquid reservoir 28, the reverse fluid microchannel 22 is connected to the lateral liquid reservoir 28. After the target fills the first lateral liquid reservoir 28, it enters the second lateral liquid reservoir 28 through the reverse fluid microchannel 22. In this way, the remaining lateral liquid reservoirs 28 are filled in turn, and the excess sample to be tested flows out from the reverse sample outlet 13 through the reverse sample outlet channel 27 ( Figure 6 The fancy arrows in the middle indicate the flow direction of the sample to be tested).

[0053] During the chip flipping process (which can also be done by oscillation), since the flow-through liquid reservoir 25 and the lateral liquid reservoir 28 are connected in the inner cavity, the probe sample in the flow-through liquid reservoir 25 and the analyte sample in the lateral liquid reservoir 28 are mixed, thereby reacting.

[0054] In the present invention, the number of reaction units, the size of the flow-through liquid reservoir 25 and the lateral liquid reservoir 28, and the number of arrays can all be adjusted.

[0055] In some embodiments of the present invention, Figure 3As shown, the flow-through reservoir 25 and the lateral reservoir 28 are both arranged in a 1×N linear array (e.g., 1×2, 1×3, 1×4). The cylindrical cavities of the N flow-through reservoirs 25 have the same radius and depth, while the N lateral reservoirs 28 have the same depth, with the cylindrical cavities increasing in radius. These are used for detecting concentration gradients of the same target. After the chip is flipped, the sample volume entering the lateral reservoir 28 from the flow-through reservoir 25 is the same. However, due to the different bottom radii of the lateral reservoirs 28, the volume varies, resulting in different reaction concentrations in each detection area. With a single sample addition, detection results for different concentration gradients of the same analyte can be simultaneously obtained.

[0056] In some embodiments of the present invention, Figure 4 As shown, the flow-through reservoir 25 and the lateral reservoir 28 are both arranged in a 1×N linear array (e.g., 1×2, 1×3, 1×4). The cylindrical cavities of the N flow-through reservoirs 25 have the same radius and depth, while the cylindrical cavities of the N lateral reservoirs 28 have the same radius and successively increasing depths, for detecting concentration gradients of the same target. After the chip is flipped, the sample volume entering the lateral reservoir 28 from the flow-through reservoir 25 is the same. Due to the different depths and thus different volumes of the lateral reservoirs 28, the reaction concentration in each detection area is different. With a single sample addition, detection results for different concentration gradients of the same analyte can be simultaneously obtained.

[0057] The flow-through liquid reservoir 25 and the lateral liquid reservoir 28 in the present invention are arranged in a 1×N linear array distribution. After the target of known concentration is injected, it is divided into multiple concentrations. The unknown concentration sample of the same target is compared with the fluorescence intensity of the multiple concentrations formed, thereby realizing quantitative analysis of the target sample, making quantitative detection more intuitive and efficient.

[0058] In some embodiments of the present invention, Figure 5As shown, the core area 2 is provided with M groups of reaction units, and the flow-through liquid reservoir 25 and the lateral liquid reservoir 28 of each group of reaction units are both distributed in a 1×N linear array, that is, the flow-through liquid reservoir 25 and the lateral liquid reservoir 28 are both distributed in an M×N rectangular array (for example, 2×3, 3×3, 4×3, 3×4, 4×4, etc.), which are used for simultaneous quantitative detection of multiple targets. The M groups of reaction units include M groups of the flow-through liquid reservoir 25 and the lateral liquid reservoir 28, which are all 1×N linear array distribution reaction units. Each group of 1×N linear array distribution reaction units has independent front and back microchannels, front and back sample inlets, and front and back sample outlets. Each group of 1×N linear array distribution reaction units introduces a probe sample. The M probe samples first introduced into the flow-through liquid reservoir 25 have independent spaces. After that, each group of 1×N linear array distribution units introduces a target into the lateral liquid reservoir 28 respectively. After the chip is flipped, the probe sample is mixed with the target in the corresponding lateral liquid reservoir 28 to form a detection array, which can detect M targets at the same time. After the chip is flipped, each group of linear arrays forms a concentration gradient of its own target, which can be used for quantitative detection.

[0059] A second aspect of the present invention provides a method for preparing the 3D printed resin-based flip chip, comprising the following steps:

[0060] Step 1: Use 3D mapping software to perform regional modeling to obtain a three-dimensional model of the upper layer 1, core area 2, and lower layer 3;

[0061] Step 2: Print the three-dimensional model of the upper layer 1, the core area 2, and the lower layer 3 using a 3D printer to obtain a printed structure;

[0062] Step 3: Cut off the base of the printed structure and use an organic solvent to remove surface impurities;

[0063] Step 4: UV curing the printed structure after impurities removal;

[0064] Step 5: Use adhesive tape to punch holes and assemble the printed structure.

[0065] In some embodiments of the present invention, the software used by the 3D printer in step 2 is Preform.

[0066] In some embodiments of the present invention, the support structure, layer thickness, and array distribution of the upper layer 1, the core area 2, and the lower layer 3 are set before printing in step 2.

[0067] In some embodiments of the present invention, the organic solvent in step 3 is isopropyl alcohol.

[0068] In some embodiments of the present invention, the UV curing in step 4 is specifically to place the printed structure after impurities removal in a UV curing box, set the temperature to 30-50° C., and the light curing time is 3-5 hours.

[0069] A third aspect of the present invention provides the use of the 3D-printed resin-based flip-type chip in detecting tumor markers, including thiol amino acids, glutathione, and cysteine.

[0070] A fourth aspect of the present invention provides a method for using the 3D printed resin-based flip chip, comprising the following steps:

[0071] S1. Sample A is injected from the front inlet 12 of the flip chip. Under pressure, sample A enters the front fluid microchannel 21 and the flow-through liquid reservoir 25 from the front inlet 12. Excess sample A is discharged from the front outlet 33 under gravity. Sample B is injected from the back inlet 32 ​​of the chip. Under pressure, sample B enters the back fluid microchannel 22 and the lateral liquid reservoir 28 from the back inlet 32.

[0072] S2. Flip the chip 180°. During the flipping process, sample A and sample B are mixed through the fluid channel between the flow-through liquid reservoir 25 and the lateral liquid reservoir 28 to react.

[0073] In some embodiments of the present invention, during the flipping and reaction process, the front sample inlet 12, the front sample outlet 33, the back sample inlet 32 ​​and the back sample outlet 13 of the chip are all sealed with adhesive tape, so that the reaction liquid is isolated from the outside during the reaction process, which is beneficial to prevent sample contamination.

[0074] In some embodiments of the present invention, the pressure in S1 is generated by an air pump or a pipette.

[0075] Below in conjunction with preferred embodiment, the specific embodiment of the present invention is described in further detail.When embodiment provides numerical range, it should be understood that, unless otherwise specified in the present invention, the two endpoints of each numerical range and any numerical value between the two endpoints can be selected.Unless otherwise defined, all technical and scientific terms used in the present invention have the same meaning as those generally understood by those skilled in the art.Except the specific method, equipment, material used in the embodiment, as those skilled in the art grasp the prior art and record of the present invention, any method, equipment and material of the prior art similar or equivalent to the method, equipment, material in the embodiments of the present invention can also be used to realize the present invention.

[0076] Example 1: Preparation process of an array of equal-depth chips with the same radius R for the core flow-through reservoir and the lateral reservoir

[0077] Import the 3D model file into the 3D printer-related software - Preform, set the width of the front fluid microchannel to 0.67mm, the radius of the cylindrical cavity of the flow-through reservoir to 2mm, the depth to 2mm, and the radius of the cylindrical cavity of the reverse lateral reservoir to 3mm. Set the arrangement to 1×3 array, the depth of the lateral reservoir to 2mm, and then connect to the 3D printer for printing. Figure 2 As shown, the structure includes 3 detection areas, 2 sample inlets and 2 sample outlets.

[0078] After printing, soak the printed structure in isopropyl alcohol solution for 3 hours to wash off excess uncured printing material; then place it in a UV post-curing box for post-curing. The curing box temperature is set to 35°C and the curing time is between 3 and 5 hours.

[0079] Example 2: Preparation process of non-uniform depth chip array with lateral reservoir radius R=3mm

[0080] Import the 3D model file into the 3D printer-related software - Preform, set the width of the front fluid microchannel to 0.67mm, the radius of the cylindrical cavity of the flow-through reservoir to 2mm, the depth to 2mm, and the radius of the cylindrical cavity of the reverse lateral reservoir to 3mm. Set the arrangement to 1×3 array, and the depth of the lateral reservoir to 2mm, 3mm, and 4mm gradients. Then connect the 3D printer for printing. Figure 3 As shown, the structure includes 3 detection areas, 2 sample inlets and 2 sample outlets.

[0081] After printing, soak the printed structure in isopropyl alcohol solution for 3 hours to wash off excess uncured printing material; then place it in a UV post-curing box for post-curing. The curing box temperature is set to 45°C and the curing time is between 3 and 5 hours.

[0082] Example 3: Preparation process of array iso-depth chips with the same radius R for flow-through reservoirs and different radius R for lateral reservoirs

[0083] The design structure diagram is as follows Figure 4 As shown, import the 3D model file into the 3D printer related software - Preform, such as Figure 5 As shown, the frontal fluid microchannel is set to a width of 0.67mm, the cylindrical cavity radius of the front flow-through reservoir is 2mm and the depth is 2.5mm, and the depth of the rear lateral reservoir is 2.5mm. The arrangement is set up in a 1×3 array, with the cylindrical cavity radius of the lateral reservoirs in a gradient of 2.5mm, 4mm, and 5.3mm. The device was then connected to a 3D printer for printing. The structure includes three detection areas, two sample inlets, and two sample outlets.

[0084] After printing, soak the printed structure in isopropyl alcohol solution for 3 hours to wash off the excess uncured printed material; then place it in a UV post-curing box for post-curing. The curing box temperature is set to 45°C and the curing time is between 3 and 5 hours. Figure 8 shown.

[0085] Example 4: The flip chip obtained in Example 3 was tested by adding a solution. Before the test, the flip chip was subjected to negative pressure treatment. The cysteine ​​detection fluorescent probe was prepared into a fluorescent probe dilution solution using ethanol: water solution (1:2, V / V). The solution was injected from the front injection port of the flip chip. The fluorescent probe dilution solution entered the front fluid microchannel from the front injection port under pressure. Since the width of the front fluid microchannel and the radius of the cylindrical cavity of the flow-through liquid reservoir were 1:3, and the inner surface of the cylindrical cavity of the flow-through liquid reservoir was hydrophilic, the fluorescent probe dilution solution was able to fill and Keep it in the flow-through reservoir, and the excess fluorescent probe is discharged from the front sample outlet under the action of gravity; then inject cysteine ​​solution from the reverse sample inlet of the flip chip, and the cysteine ​​solution enters the reverse fluid microchannel and the lateral liquid reservoir from the reverse sample inlet under the action of pressure; the front sample inlet, front sample outlet, reverse sample inlet and reverse sample outlet of the chip are all sealed with adhesive tape; flip the chip 180 degrees, and the fluorescent probe and cysteine ​​solution are mixed through the fluid channel between the flow-through reservoir and the lateral liquid reservoir to obtain three concentration gradients of fluorescent probe and target mixed solutions. The actual picture of the obtained flip chip after adding fluorescent probe and detecting target is as follows Figure 9 Part of the solution filled the cavity, and fluorescence microscopy was used to analyze the color development results of various fluorescence channels. RGB color blocks were used for regional analysis to obtain the concentration gradient of cysteine.

[0086] The above description is merely a preferred embodiment of the present invention and is not intended to limit the scope of the present invention. Various modifications are possible. Any simple, equivalent changes and modifications made in accordance with the claims and description of the present invention are within the scope of protection of the patent claims. Anything not fully described in this invention is conventional technology.

Claims

1. A 3D printed resin-based flip chip, characterized in that: The flip chip comprises an upper layer (1), a core area (2), and a lower layer (3) bonded sequentially from top to bottom; the core area (2) is provided with at least one group of reaction units, each group of reaction units comprising a front fluid microchannel (21) and a back fluid microchannel (22); The front fluid microchannel (21) is provided with a plurality of flow-through liquid reservoirs (25), the front fluid microchannel (21) horizontally passes through the central area of ​​the flow-through liquid reservoir (25), and the inlet and outlet of the front fluid microchannel (21) are symmetrically distributed relative to the flow-through liquid reservoir (25); a plurality of lateral liquid reservoirs (28) are provided on one side of the reverse fluid microchannel (22), the reverse fluid microchannel (22) does not pass through the central area of ​​the lateral liquid reservoir (28), and the inlet and outlet of the reverse fluid microchannel (22) are located beside the lateral liquid reservoir (28); The flow-through liquid reservoir (25) and the lateral liquid reservoir (28) are two cylinders connected in an integral manner from top to bottom. The inner cavity of the flow-through liquid reservoir (25) and the inner cavity of the lateral liquid reservoir (28) are communicated through a fluid channel (29). The fluid channel (29) is a through hole provided at the center of the top surface of the lateral liquid reservoir (28). The ratio of the width of the front fluid microchannel (21) to the radius of the cylindrical cavity of the flow-through liquid reservoir (25) is 1: :2~4, the inner surface of the cylindrical cavity of the flow-through liquid reservoir (25) is hydrophilic, so that the sample 1 is kept in the flow-through liquid reservoir (25) in a non-inverted state; the width of the reverse fluid microchannel (22) and the radius of the cylindrical cavity of the lateral liquid reservoir (28) are 1:2~4, and the inner surface of the cylindrical cavity of the lateral liquid reservoir (28) is hydrophilic, so that the sample 2 is kept in the lateral liquid reservoir (28) in a non-inverted state; The upper layer (1) is provided with a front sampling groove (11) communicating with the front fluid microchannel (21), a front sampling port (12), and a rear sampling port (13) communicating with the rear fluid microchannel (22); the lower layer (3) is provided with a rear sampling groove (31) communicating with the rear fluid microchannel (22), a rear sampling port (32), and a front sampling port (33) communicating with the front fluid microchannel (21); During injection, sample 1 and sample 2 are retained independently in the flow-through liquid reservoir (25) and the lateral liquid reservoir (28), respectively, and the two are mixed through the fluid channel (29) by flipping the chip.

2. The 3D printed resin-based flip chip according to claim 1, wherein: The inner diameter of the cylindrical cavity of the flow-through liquid storage tank (25) is smaller than the inner diameter of the cylindrical cavity of the lateral liquid storage tank (28).

3. The 3D printed resin-based flip chip according to claim 1, wherein: The front fluid microchannel (21) and the reverse fluid microchannel (22) of the core area (2) are horizontally distributed; one end of the front fluid microchannel (21) is provided with a vertically upward front sample inlet channel (23), and the other end is provided with a vertically downward front sample outlet channel (24); one end of the reverse fluid microchannel (22) is provided with a vertically downward reverse sample inlet channel (26), and the other end is provided with a vertically upward reverse sample outlet channel (27); the front sample inlet groove (11) is located below the upper layer (1) and is perpendicular to the horizontal direction of the front sample inlet channel (23); the reverse sample inlet groove (31) is located below the lower layer (3) and is perpendicular to the horizontal direction of the reverse sample inlet channel (26).

4. The 3D printed resin-based flip chip according to claim 1, wherein: Also includes any of the following features: 1) The flow-through liquid reservoir (25) and the lateral liquid reservoir (28) are both distributed in a 1×N linear array, the cylindrical cavity radius and depth of the N flow-through liquid reservoirs (25) are the same, the depth of the N lateral liquid reservoirs (28) is the same, and the cylindrical cavity radius increases successively, so as to form a concentration gradient by injecting the same target substance once; 2) The flow-through liquid reservoir (25) and the lateral liquid reservoir (28) are both distributed in a 1×N linear array, the cylindrical cavity radius and depth of the N flow-through liquid reservoirs are the same, and the cylindrical cavity radius of the N lateral liquid reservoirs is the same, and the depth increases successively, so as to form a concentration gradient by injecting the same target object once.

5. The 3D printed resin-based flip chip according to any one of claims 4, wherein: The core area (2) is provided with M groups of reaction units, and the flow-through liquid storage tank (25) and the lateral liquid storage tank (28) of each group of reaction units are both distributed in a 1×N linear array for simultaneous quantitative detection of multiple targets.

6. A method for preparing a 3D printed resin-based flip chip according to any one of claims 1 to 5, characterized in that: The following steps are involved: Step 1: Use 3D mapping software to perform regional modeling to obtain three-dimensional models of the upper layer, core area, and lower layer; Step 2: Use a 3D printer to print a three-dimensional model of the upper layer, core area, and lower layer to obtain a printed structure; Step 3: Cut off the base of the printed structure and use an organic solvent to remove surface impurities; Step 4: UV curing the printed structure after impurities removal; Step 5: Use adhesive tape to punch holes and assemble the printed structure.

7. The method for preparing a 3D printed resin-based flip chip according to claim 6, wherein: Also includes one or more of the following characteristics: i) The software used by the 3D printer in step 2 is Preform; ii) Setting the support structure, layer thickness, and array distribution of the upper layer, core area, and lower layer before printing in step 2; iii) the organic solvent in step 3 is isopropanol; ⅵ) The UV curing in step 4 specifically includes placing the printed structure after impurity removal in a UV curing box, setting the temperature to 30-50°C, and curing for 3-5 hours.

8. Use of the 3D-printed resin-based flip chip according to any one of claims 1 to 5 in tumor marker detection.

9. A method for using a 3D printed resin-based flip chip according to any one of claims 1 to 5, comprising the following steps: S1. Inject sample A from the front inlet of the flip chip. Under pressure, sample A enters the front fluid microchannel and flow-through reservoir from the front inlet. Excess sample A is discharged from the front outlet under gravity. Inject sample B from the inlet on the back of the chip. Under pressure, sample B enters the reverse fluid microchannel and the lateral liquid reservoir from the inlet on the back. S2. Flip the chip 180°. During the flipping process, sample A and sample B are mixed through the fluid channel between the flow-through liquid reservoir and the lateral liquid reservoir to react.

10. The method of use according to claim 9, wherein: Also includes one or more of the following characteristics: 1) During the flipping and reaction process, the front inlet, front outlet, back inlet, and back outlet of the chip are sealed with adhesive tape; 2) Before the sample is injected in S1, the chip is subjected to negative pressure treatment, and the pressure effect is generated by the pressure difference.

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