Microfluidic organoid drug screening chip

Abstract

The utility model discloses a microfluidic organoid drug screening chip, which belongs to the field of microfluidic chips, and comprises M drug channels arranged in a circular array form around the same centrifugal shaft, and one or more sample introduction channels extending from a near-shaft end to a far-shaft end along the circumferential direction in a spiral line form around the centrifugal shaft, the medicine channel extends from the near-shaft end to the far-shaft end in the radial direction in a broken line mode, a medicine inlet is formed in the head end of the medicine channel, the tail end of the medicine channel is connected with the first waste liquid cavity, and C medicine quantifying cavities are formed in the outer side of the medicine channel; the head end of the sample introduction channel is provided with a sample adding chamber, the tail end of the sample introduction channel is connected with a second waste liquid chamber, and N sample quantification chambers in one-to-one correspondence with the M * C medicine quantification chambers are arranged on the outer side of the sample introduction channel; the sample quantification chamber is arranged at the downstream of the corresponding medicine quantification chamber, and the sample quantification chamber and the medicine quantification chamber are connected through a micro-channel. The form of the medicine channel and the form of the sample injection channel are interchangeable. According to the organ-like drug screening device, high-flux, low-cost, integrated and automatic organ-like drug screening can be realized.

Description

Technical Field

[0001] The utility model belongs to the field of microfluidic chips, and more specifically, relates to a microfluidic organoid drug screening chip. Background Art

[0002] As a bridge between traditional 2D culture and animal models, organoids have many advantages, such as a physiological environment closer to in vivo cells, a stable genome, and a short culture cycle, which can provide experimental operability. At present, organoids have shown great potential in clinical tumor drug screening and drug development. However, in the face of the high-throughput, multiple drug combinations, short-term high efficiency and low cost requirements of drug screening, the traditional detection method based on microplate spotting has shown many inconveniences in organoid drug screening applications.

[0003] As an emerging science and technology, microfluidics has been applied to many fields such as chemistry, biology, engineering and physics. It has strong interdisciplinary nature and has made breakthroughs in the precise control of time, space and analysis objects, and can solve many key problems in life analysis. Microfluidics can integrate detection experiments that could only be completed in the laboratory into a small chip, which not only saves consumables and time costs, but more importantly, it can integrate multiple detection technologies into one to improve detection efficiency.

[0004] Nowadays, microfluidic technology has been used and promoted in the field of bacterial drug sensitivity detection, but its application in the field of organoids is still in its infancy. The currently used off-chip perfusion technology makes the chip structure complex and the cost high. How to achieve high-throughput, low-cost, integrated and automated organoid drug screening is still a problem that needs to be solved urgently. Utility Model Content

[0005] In view of the defects of the prior art and the need for improvement, the utility model provides a microfluidic organoid drug screening chip, the purpose of which is to improve the structure of the microfluidic chip so that it can achieve high-throughput, low-cost, integrated and automated organoid drug screening.

[0006] To achieve the above-mentioned object, according to one aspect of the utility model, there is provided a microfluidic organoid drug screening chip, comprising: M drug channels arranged in a circular array around the same centrifugal axis, and one or more injection channels extending from a proximal end to a distal end in a spiral line along a circumferential direction around the centrifugal axis;

[0007] The drug channel extends radially from the proximal end to the distal end in a broken line form, a drug inlet is arranged at the head end, a terminal end is connected to the first waste liquid chamber, and C drug quantitative chambers are arranged outside the drug channel; the first waste liquid chamber is connected to the atmosphere;

[0008] The sampling channel has a sample loading chamber at its head end, a second waste liquid chamber at its tail end, and N sample quantitative chambers at its outer side; the sample loading chamber is provided with a sample loading port, and the second waste liquid chamber is connected to an air hole connected to the atmosphere through an airway;

[0009] M, N, and C are all positive integers, and N=M×C, and the N sample quantification chambers correspond one to one with the M×C drug quantification chambers; the sample quantification chamber is arranged downstream of the corresponding drug quantification chamber, and the top of the sample quantification chamber is connected to the bottom of the corresponding drug quantification chamber through a microchannel.

[0010] Furthermore, the microfluidic organoid drug screening chip provided by the utility model comprises, from top to bottom, a cover plate, a drug layer, a microchannel layer, an organoid layer and a bottom plate;

[0011] The drug channel is arranged in the drug layer;

[0012] The microchannel is arranged in the microchannel layer;

[0013] The injection channel is set in the organoid layer;

[0014] The drug inlet and the sample adding port are both arranged on the cover plate; the drug inlet is matched with the head end of each drug channel, and the sample adding port is matched with the sample adding chamber;

[0015] The drug layer is also provided with an airway connected to the first waste liquid chamber, the organoid layer is also provided with an airway connected to the second waste liquid chamber, and the cover plate is also provided with air holes connected to the atmosphere, and the air holes are matched with the airway ends of the drug layer and the organoid layer.

[0016] Furthermore, the volume of the drug quantitative chamber on the same drug channel gradually increases or decreases.

[0017] Furthermore, the drug quantitative chamber and the corresponding sample quantitative chamber on the same drug channel are located in the same radial direction.

[0018] Furthermore, the microchannel is serpentine-shaped.

[0019] According to another aspect of the utility model, a microfluidic organoid drug screening chip is provided, comprising: M injection channels arranged in a circular array around the same centrifugal axis, and one or more drug channels extending from a proximal end to a distal end in a spiral line along a circumferential direction around the centrifugal axis;

[0020] The sampling channel extends radially from the proximal end to the distal end in a broken line form, a sampling port is arranged at the head end, the end is connected to the first waste liquid chamber, and C sample quantitative chambers are arranged outside the channel; the first waste liquid chamber is connected to the atmosphere;

[0021] The drug channel has a drug chamber at its head end, a second waste liquid chamber at its tail end, and N drug quantitative chambers at its outer side; the drug chamber is provided with a drug inlet, and the second waste liquid chamber is connected to an air hole connected to the atmosphere through an airway;

[0022] M, N, and C are all positive integers, and N=M×C, and the N drug quantitative chambers correspond one to one with the M×C sample quantitative chambers; the sample quantitative chamber is arranged downstream of the corresponding drug quantitative chamber, and the top of the sample quantitative chamber is connected to the bottom of the corresponding drug quantitative chamber through a microchannel.

[0023] Furthermore, the microfluidic organoid drug screening chip provided by the utility model comprises, from top to bottom, a cover plate, a drug layer, a microchannel layer, an organoid layer and a bottom plate;

[0024] The drug channel is arranged in the drug layer;

[0025] The microchannel is arranged in the microchannel layer;

[0026] The injection channel is set in the organoid layer;

[0027] The drug inlet and the sample adding port are both arranged on the cover plate; the drug inlet is matched with the head end of each drug channel, and the sample adding port is matched with the sample adding chamber;

[0028] The organoid layer is also provided with an airway connected to the first waste liquid chamber, the drug layer is also provided with an airway connected to the second waste liquid chamber, and the cover plate is also provided with air holes connected to the atmosphere, and the air holes are matched with the airway ends of the drug layer and the organoid layer.

[0029] Furthermore, the volume of the sample quantification chamber on the same injection channel gradually increases or decreases.

[0030] Furthermore, the sample quantitative chamber and the corresponding drug quantitative chamber on the same sample channel are located in the same radial direction.

[0031] Furthermore, the microchannel is serpentine-shaped.

[0032] In general, the above technical solutions conceived by the present invention can achieve the following beneficial effects:

[0033] (1) The microfluidic organoid drug screening chip provided by the utility model has a sampling channel and a drug channel, one of which extends from the proximal end to the distal end in a spiral line along the circumferential direction around the centrifugal axis, and the other channel is arranged in a circular array around the centrifugal axis, and extends from the proximal end to the distal end in a broken line along the radial direction, so that the two types of channels are arranged in a radial and tangential cross-combination layout. At the same time, the sampling channel and the drug channel are provided with a one-to-one corresponding and interconnected sample quantification chamber and drug quantification chamber. Combined with the cross-combination channel layout, a variety of different organoid and drug combination reactions can be achieved at the same time, and finally high-throughput, homogenized, and automated organoid drug screening can be achieved; in addition, the entire process of organoid drug screening is integrated into the chip, with high integration, and the entire process only needs to be controlled by centrifugation, and the cost is low. In general, the utility model can achieve high-throughput, low-cost, integrated, and automated organoid drug screening.

[0034] (2) The microfluidic organoid drug screening chip provided by the utility model comprises a drug layer, a microchannel layer and an organoid layer in the adjacent upper, middle and lower layers respectively, and the upper and lower layers are connected only by a microchannel, forming a stepped structure of "drug quantification chamber - microchannel - sample quantification chamber", which can conveniently realize the primary quantification of drugs and samples, as well as the secondary quantification of the same volume after the drugs and samples are mixed.

[0035] (3) In the microfluidic organoid drug screening chip provided by the utility model, the volume of the quantitative chamber on the radially extending injection channel or drug channel gradually increases or decreases, so that different concentration gradients can be formed on the same channel, thereby realizing a combination of organoids and drugs of the same type but different concentrations, and achieving a richer combination reaction.

[0036] (4) In the microfluidic organoid drug screening chip provided by the utility model, the corresponding drug quantification chamber and sample quantification chamber on the same radial extension channel are located in the same radial direction, that is, the projections of the corresponding drug quantification chamber and sample quantification chamber on the same radial extension channel on the chip surface are located on the same radial straight line, ensuring that each drug of each concentration gradient corresponds to the organoid one-to-one. At the same time, the same straight line is in the same direction as the centrifugal force, making drug injection more labor-saving and accurate.

[0037] (5) The microfluidic tailpipe drug screening chip provided by the utility model has a serpentine-shaped microchannel for connecting the corresponding sample quantification chamber and the drug quantification chamber, thereby increasing the fluid resistance and facilitating the control of the connectivity between the chambers. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 Schematic diagram of the microfluidic organoid drug screening chip provided in Example 1 of the utility model;

[0039] Figure 2 A schematic diagram of the hierarchical design of the microfluidic organoid drug screening chip provided in Example 1 of the utility model;

[0040] Figure 3 Schematic diagram of a microfluidic organoid drug screening chip provided in Example 2 of the utility model;

[0041] Figure 4 A schematic diagram of the hierarchical design of a microfluidic organoid drug screening chip provided in Example 2 of the utility model;

[0042] Throughout the drawings, the same reference numerals are used to denote the same elements or structures, wherein:

[0043] 11-first waste liquid chamber, 12-second waste liquid chamber, 13-microchannel, 14-sample addition chamber, 15-drug channel, 16-injection channel, 17-drug quantitative chamber, 18-sample quantitative chamber;

[0044] 21 - first waste liquid chamber, 22 - second waste liquid chamber, 23 - microchannel, 24 - drug chamber, 25 - drug channel, 26 - injection channel, 27 - drug quantitative chamber, 28 - sample quantitative chamber. DETAILED DESCRIPTION

[0045] In order to make the purpose, technical solution and advantages of the utility model more clear, the utility model is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the utility model and are not used to limit the utility model. In addition, the technical features involved in each embodiment of the utility model described below can be combined with each other as long as they do not conflict with each other.

[0046] In the present utility model, the terms "first", "second", etc. (if any) in the present utility model and the accompanying drawings are used to distinguish similar objects, and are not necessarily used to describe a specific order or sequence. In addition, in the description of the present utility model, the terms "upper", "lower", "left", "right", "inner", "outer", etc. indicating directions or positional relationships are based on the directions or positional relationships shown in the accompanying drawings. This is only for the convenience of description, and does not indicate or imply that the device or element must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation of the present utility model. In addition, it should be noted that in the description of the present utility model, unless otherwise clearly specified and limited, the terms "installed", "connected", and "connected" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be directly connected, or indirectly connected through an intermediate medium, or it can be a connection between the two elements. For those skilled in the art, the specific meanings of the above terms in the present utility model can be understood according to the specific circumstances.

[0047] It is easy to understand that terms such as "inside", "outside", "upstream", "downstream", "high", "low", "proximal end" and "distal end" in the centrifugal chip are all relative to the distance from the centrifugal axis. Specifically, the side closer to the centrifugal axis is the "inside", and the side farther from the centrifugal axis is the "outside"; the point closer to the centrifugal axis is located "upstream", and the point farther from the centrifugal axis is located "downstream"; the point closer to the centrifugal axis is the higher point, and the point farther from the centrifugal axis is the lower point; the end closer to the centrifugal axis is the proximal end, and the end farther from the centrifugal axis is the distal end.

[0048] In order to achieve high-throughput, low-cost, integrated and automated organoid drug screening, the utility model provides a microfluidic organoid drug screening chip, the overall idea of ​​which is to improve the chip structure so that the sampling flow channel and the drug flow are arranged in a cross-combination layout, thereby achieving a more complex analysis combination, improving the detection throughput without increasing the cost, and integrating the entire process of organoid drug screening into the chip to achieve automated detection.

[0049] The following are examples.

[0050] Embodiment 1:

[0051] A microfluidic organoid drug screening chip, such as Figure 1 As shown, it includes: 10 drug channels 15 arranged in a circular array around the same centrifugal axis, and an injection channel 16 extending from the proximal end to the distal end in a spiral line along the circumferential direction around the centrifugal axis;

[0052] The drug channel 15 extends radially from the proximal end to the distal end in a broken line form, a drug inlet is provided at its head end, and its tail end is connected to the first waste liquid chamber 11, and three drug quantitative chambers 17 are provided outside thereof; the first waste liquid chamber 11 is connected to the air hole connected to the atmosphere through the airway; a total of 30 drug quantitative chambers are provided on the 10 drug channels;

[0053] The sampling channel 16 has a sample adding chamber 14 at its head end, and its end is connected to the second waste liquid chamber 12, and 30 sample quantitative chambers 18 are arranged outside the sample adding chamber 14; the sample adding chamber 14 is provided with a sample adding port, and the second waste liquid chamber 12 is connected to the air hole connected to the atmosphere through the airway;

[0054] The 30 sample quantification chambers 18 correspond one to one with the 30 drug quantification chambers 17 ; the sample quantification chamber 18 is arranged downstream of the corresponding drug quantification chamber 17 , and the top of the sample quantification chamber 18 is connected to the bottom of the corresponding drug quantification chamber 17 through the microchannel 13 .

[0055] like Figure 2 As shown, the microfluidic organoid drug screening chip provided in this embodiment includes multiple chip layers, specifically, from top to bottom, it includes: a cover plate, a drug layer, a microchannel layer, an organoid layer and a bottom plate, respectively. Figure 2 As shown in (a), (b), (c), (d) and (e);

[0056] The drug channel is arranged in the drug layer;

[0057] The microchannel is arranged in the microchannel layer;

[0058] The injection channel is set in the organoid layer;

[0059] The drug inlet, the sample addition port and the air holes are all arranged on the cover plate; the drug inlet is matched with the head end of each drug channel, the sample addition port is matched with the sample addition chamber, and the air hole is matched with the airway end of the drug layer and the organoid layer;

[0060] To facilitate chip assembly, Figure 2 As shown, each chip layer is provided with a plurality of positioning holes, one of which is a central positioning hole, which coincides with the centrifugal axis and is used to fix the chip on the centrifuge.

[0061] In this embodiment, the chip processing material can be a transparent material such as PMMA, PDMS, etc., which can be directly used for detection and analysis by optical instruments.

[0062] In practical applications, after adding the organoid sample suspension to the sample loading chamber, centrifugation allows the organoid sample suspension to enter the sample quantification chamber through the injection channel, and the excess organoid sample suspension will enter the second waste liquid chamber; adding drugs to the drug inlet and centrifuging allows the drugs to enter the drug quantification chamber through the drug channel, and the excess drugs will enter the first waste liquid chamber; due to the small space of the microchannel, the tortuous flow channel, and the large fluid resistance, the drug solution will stay in the drug quantification chamber and cannot break through the hydrophobic resistance to enter the lower sample quantification chamber. After a certain period of time, the liquid in the sample quantification chamber will gradually evaporate, thereby reserving a certain space in the sample quantification chamber. At this time, with the help of high-speed centrifugation, the drug solution breaks through the microchannel and enters the sample quantification chamber to mix evenly with the organoid suspension in the chamber to complete the second quantification. After that, the drug and organoid can be incubated to complete the organoid screening test.

[0063] In this embodiment, a single chip is provided with 10 drug injection channels, and each drug channel is provided with 3 drug quantification chambers of different volumes, that is, a maximum of 10 different drugs can be combined with the same organoid, and each drug can be combined with 3 different concentrations. Therefore, this embodiment can achieve 30 different combinations.

[0064] Since the injection channel is spiral-shaped, injection is more convenient.

[0065] In practical applications, different types of drugs can be added to different drug channels, or different concentrations of drugs of the same type can be added. In order to facilitate the detection of different concentrations of the same type of drug, as a preferred embodiment, in this embodiment, the volume of the drug quantitative chamber on the same drug channel gradually increases. It is easy to understand that in some other embodiments, the volume of the drug quantitative chamber on the same drug channel can also be set to gradually increase, and the gradient concentration of the same drug can also be achieved.

[0066] In order to facilitate the distinction of the detection results corresponding to different drug channels in actual detection, as a preferred implementation, in this embodiment, the drug quantification chamber and the corresponding sample quantification chamber on the same drug channel are located in the same radial direction.

[0067] like Figure 1 and Figure 2 As shown, as a preferred embodiment, in this embodiment, the microchannel is serpentine, thereby increasing the fluid resistance and facilitating the control of the communication relationship between the drug quantitative chamber and the sample quantitative chamber. In addition, in this embodiment, a valve is also provided at the head end of the microchannel, which can be a hydrophobic valve, a sacrificial valve or a soluble membrane valve.

[0068] In this embodiment, the second waste liquid chamber connected to the spiral injection channel is annularly arranged around the outermost side of the chip, thereby saving chip space.

[0069] It should be noted that, in this embodiment, the number of injection channels and drug channels, as well as the number of quantitative chambers on each channel, are merely exemplary descriptions and should not be construed as the sole limitation of the present invention. In practical applications, they can be flexibly set according to actual organoid drug screening requirements.

[0070] The working principle of the microfluidic organoid drug screening chip provided in this embodiment is explained below in conjunction with a specific organoid drug screening example. The detection process is as follows:

[0071] S1: slowly add the organoid suspension into the sample loading chamber, and then push or centrifuge at a low speed to evenly distribute the organoid suspension to each sample quantitative chamber, and the excess sample solution enters the second quantitative chamber;

[0072] S2: slowly add the drug solution through the injection port of each drug injection channel, and push the solution to distribute the drug concentration gradient, and the drug solution enters the drug quantitative chambers of different volumes to complete the first quantification; due to the small space of the microchannel, the tortuous flow channel, and the large fluid resistance, the drug solution will stay in the drug quantitative chamber and cannot break through the hydrophobic resistance to enter the lower sample quantitative chamber; it is easy to understand that the 10 drug channels here can select ten different drugs or drug combinations according to the needs of drug screening, or can select fewer drug types and different concentrations of drug concentrations;

[0073] S3: After a certain period of time, the liquid in the sample quantification chamber will gradually evaporate, thereby reserving a certain space in the sample quantification chamber. At this time, the centrifuge speed is increased. Under high-speed centrifugation, the drug solution breaks through the microchannel and enters the sample quantification chamber to mix evenly with the organoid suspension in the chamber to complete the second quantification;

[0074] S4: After the drug and organoids are incubated for a period of time, the organoid activity detection reagent (such as calcein-propidium iodide PBS solution) is added again through the injection channel in the same way as S1, and the excess solution enters the second waste liquid chamber.

[0075] S5: After the detection reagent and organoid react completely, the test results are read.

[0076] In general, the microfluidic organoid drug screening chip provided in this embodiment has the following characteristics:

[0077] (1) It includes multiple sample quantification chambers, and each drug injection channel includes multiple drug quantification chambers. It can realize drug screening and drug sensitivity detection for different types of drugs. For the same type of drug, due to the different volumes of the drug quantification chambers, a concentration gradient can be automatically generated, thereby quickly detecting the IC50 (half maximal inhibitory concentration) value, greatly reducing the time and cost of manual or mechanical drug gradient configuration, and quickly and accurately realizing the screening of single or multiple drug combinations. In addition, the reagent consumption of the chip is in the nanoliter level, which greatly reduces the cost of use.

[0078] (2) It includes a tangentially arranged organoid sampling channel and a radially arranged drug sampling channel, forming an XY cross combination layout. At the same time, the sample flow channel and the drug flow channel are provided with a one-to-one corresponding and interconnected sample quantification chamber and drug quantification chamber. Through this flow channel layout, the utility model can control the mixing between the sample and the drug from the tangential and radial dimensions by centrifugation, and realize the primary quantification of different volumes of drugs and the secondary quantification of the same volume after the drug and sample are mixed. In this embodiment, the same chip provides at least dozens of quantitative chambers for simultaneous reaction, and finally realizes high-throughput, homogenized, and automated drug screening.

[0079] Embodiment 2:

[0080] A microfluidic organoid drug screening chip. This embodiment is similar to the above-mentioned embodiment 1, except that, in this embodiment, the design of the injection channel and the drug channel is swapped, the drug channel extends in a spiral shape along the circumferential direction, and the injection channel extends in a broken line shape along the radial direction.

[0081] Specific as Figure 3 As shown, in this embodiment, it includes: 10 injection channels 26 arranged in a circular array around the same centrifugal axis, and 1 drug channel 25 extending from the proximal end to the distal end in a spiral line along the circumferential direction around the centrifugal axis;

[0082] The injection channel 26 extends radially from the proximal end to the distal end in a broken line form, a sample injection port is provided at its head end, and its end is connected to the first waste liquid chamber 21, and three sample quantitative chambers 28 are provided outside thereof; the first waste liquid chamber 21 is connected to the air hole connected to the atmosphere through the airway; a total of 30 sample quantitative chambers 28 are provided on the 10 injection channels 26;

[0083] The drug channel 25 has a drug chamber 24 at its head end, and its tail end is connected to the second waste liquid chamber 22, and 30 drug quantitative chambers 27 are arranged outside the drug channel; the drug chamber 24 is provided with a drug inlet, and the second waste liquid chamber 22 is connected to the air hole connected to the atmosphere through the airway;

[0084] The 30 drug quantitative chambers 27 correspond to the 30 sample quantitative chambers 28 one by one; the sample quantitative chamber 28 is arranged downstream of the corresponding drug quantitative chamber 27, and the top of the sample quantitative chamber 28 is connected to the bottom of the corresponding drug quantitative chamber 27 through the microchannel 23.

[0085] like Figure 4 As shown, the microfluidic organoid drug screening chip provided in this embodiment includes, from top to bottom, a cover plate, a drug layer, a microchannel layer, an organoid layer and a bottom plate, respectively. Figure 4 As shown in (a), (b), (c), (d) and (e);

[0086] The drug channel is arranged in the drug layer;

[0087] The microchannel is arranged in the microchannel layer;

[0088] The injection channel is set in the organoid layer;

[0089] The drug inlet, the sample addition port and the air holes are all arranged on the cover plate; the drug inlet is matched with the head end of each drug channel, the sample addition port is matched with the sample addition chamber, and the air hole is matched with the airway end of the drug layer and the organoid layer;

[0090] To facilitate chip assembly, Figure 4 As shown, each chip layer is provided with a plurality of positioning holes, one of which is a central positioning hole, which coincides with the centrifugal axis and is used to fix the chip on the centrifuge.

[0091] In this embodiment, the chip processing material can be a transparent material such as PMMA, PDMS, etc., which can be directly used for detection and analysis by optical instruments.

[0092] The microfluidic organoid drug screening chip provided in this embodiment has a working principle and working method similar to the above-mentioned embodiment 1. Specifically, after adding the organoid sample suspension to the sample addition port, the organoid sample suspension can enter the sample quantification chamber through the injection channel through centrifugation, and the excess organoid sample suspension will enter the first waste liquid chamber; after adding the drug to the drug chamber and centrifuging, the drug can enter the drug quantification chamber through the drug channel, and the excess drug will enter the second waste liquid chamber; due to the narrow space of the microchannel, the tortuous flow channel, and the large fluid resistance, the drug solution will stay in the drug quantification chamber and cannot break through the hydrophobic resistance to enter the lower sample quantification chamber. After a certain period of time, the liquid in the sample quantification chamber will gradually evaporate, thereby reserving a certain space in the sample quantification chamber. At this time, with the help of high-speed centrifugation, the drug solution breaks through the microchannel and enters the sample quantification chamber to mix evenly with the organoid suspension in the chamber to complete the second quantification, and then the drug and organoid can be incubated to complete the organoid screening test.

[0093] In this embodiment, a single chip is provided with 10 drug injection channels, and each drug channel is provided with 3 drug quantification chambers of different volumes, that is, a maximum of 10 different drugs can be combined with the same organoid, and each drug can be combined with 3 different concentrations. Therefore, this embodiment can achieve 30 different combinations.

[0094] Since the injection channel is spiral-shaped, injection is more convenient.

[0095] In practical applications, different types of drugs can be added to different drug channels, or different concentrations of drugs of the same type can be added. In order to facilitate the detection of different concentrations of the same type of drug, as a preferred embodiment, in this embodiment, the volume of the drug quantitative chamber on the same drug channel gradually increases. It is easy to understand that in some other embodiments, the volume of the drug quantitative chamber on the same drug channel can also be set to gradually increase, and the gradient concentration of the same drug can also be achieved.

[0096] In order to facilitate the distinction of the detection results corresponding to different drug channels in actual detection, as a preferred implementation, in this embodiment, the drug quantification chamber and the corresponding sample quantification chamber on the same drug channel are located in the same radial direction.

[0097] like Figure 3 and Figure 4 As shown, as a preferred embodiment, in this embodiment, the microchannel is serpentine, thereby increasing the fluid resistance and facilitating the control of the communication relationship between the drug quantitative chamber and the sample quantitative chamber. In addition, in this embodiment, a valve is also provided at the head end of the microchannel, which can be a hydrophobic valve, a sacrificial valve or a soluble membrane valve.

[0098] In this embodiment, the second waste liquid chamber connected to the spiral injection channel is annularly arranged around the outermost side of the chip, thereby saving chip space.

[0099] It should also be noted that, in this embodiment, the number of injection channels and drug channels, as well as the number of quantitative chambers on each channel, are merely exemplary descriptions and should not be construed as the sole limitation of the present invention. In practical applications, they can be flexibly set according to actual organoid drug screening requirements.

[0100] Similar to the above-mentioned embodiment 1, the microfluidic organoid drug screening chip provided in this embodiment has the following characteristics:

[0101] (1) It includes multiple sample quantification chambers, and each drug injection channel includes multiple drug quantification chambers. It can realize drug screening and drug sensitivity detection for different types of drugs. For the same type of drug, due to the different volumes of the drug quantification chambers, a concentration gradient can be automatically generated, thereby quickly detecting the IC50 (half maximal inhibitory concentration) value, greatly reducing the time and cost of manual or mechanical drug gradient configuration, and quickly and accurately realizing the screening of single or multiple drug combinations. In addition, the reagent consumption of the chip is in the nanoliter level, which greatly reduces the cost of use.

[0102] (2) It includes a tangentially arranged organoid sampling channel and a radially arranged drug sampling channel, forming an XY cross combination layout. At the same time, the sample flow channel and the drug flow channel are provided with a one-to-one corresponding and interconnected sample quantification chamber and drug quantification chamber. Through this flow channel layout, the utility model can control the mixing between the sample and the drug from the tangential and radial dimensions by centrifugation, and realize the primary quantification of different volumes of drugs and the secondary quantification of the same volume after the drug and sample are mixed. In this embodiment, the same chip provides at least dozens of quantitative chambers for simultaneous reaction, and finally realizes high-throughput, homogenized, and automated drug screening.

[0103] Combining Example 1 and Example 2, it can be seen that the utility model provides a microfluidic organoid high-throughput drug screening chip with simple driving force, convenient operation, high integration, low cost, high throughput, rapid and accurate, which can be used for rapid detection and screening of samples and multiple drugs or drug combinations. In addition, it can realize the reaction of multiple drugs with the same organoid sample, and it can also realize the reaction of multiple organoid samples with the same drug, which can be suitable for different application scenarios.

[0104] It is easy for those skilled in the art to understand that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A microfluidic organoid drug screening chip, characterized in that: include: M drug channels arranged in a circular array around the same centrifugal axis, and one or more injection channels extending in a spiral line along the circumferential direction from the proximal end to the distal end around the centrifugal axis; The drug channel extends radially from the proximal end to the distal end in a broken line form, a drug inlet is arranged at the head end, a terminal end is connected to the first waste liquid chamber, and C drug quantitative chambers are arranged outside the drug channel; the first waste liquid chamber is connected to the atmosphere; The sampling channel has a sample loading chamber at its head end, a second waste liquid chamber at its tail end, and N sample quantitative chambers at its outer side; the sample loading chamber is provided with a sample loading port, and the second waste liquid chamber is connected to an air hole connected to the atmosphere through an airway; M, N, and C are all positive integers, and N=M×C, and the N sample quantification chambers correspond one to one with the M×C drug quantification chambers; the sample quantification chamber is arranged downstream of the corresponding drug quantification chamber, and the top of the sample quantification chamber is connected to the bottom of the corresponding drug quantification chamber through a microchannel.

2. The microfluidic organoid drug screening chip according to claim 1, characterized in that: From top to bottom, it includes: cover plate, drug layer, microchannel layer, organoid layer and bottom plate; The drug channel is arranged in the drug layer; The microchannel is disposed in the microchannel layer; The injection channel is arranged in the organoid layer; The drug inlet and the sample adding port are both arranged on the cover plate; the drug inlet is matched with the head end of each drug channel, and the sample adding port is matched with the sample adding chamber; The drug layer is also provided with an airway connected to the first waste liquid chamber, the organoid layer is also provided with an airway connected to the second waste liquid chamber, and the cover plate is also provided with air holes connected to the atmosphere, and the air holes are matched with the airway ends of the drug layer and the organoid layer.

3. The microfluidic organoid drug screening chip according to claim 1 or 2, characterized in that: The volume of the drug quantitative chamber on the same drug channel gradually increases or decreases.

4. The microfluidic organoid drug screening chip according to claim 1 or 2, characterized in that: The drug quantitative chamber and the corresponding sample quantitative chamber on the same drug channel are located in the same radial direction.

5. The microfluidic organoid drug screening chip according to claim 1 or 2, characterized in that: The microchannel is in a serpentine shape.

6. A microfluidic organoid drug screening chip, characterized in that: include: M injection channels arranged in a circular array around the same centrifugal axis, and one or more drug channels extending in a spiral line along the circumferential direction from the proximal end to the distal end around the centrifugal axis; The injection channel extends radially from the proximal end to the distal end in a broken line form, a sample injection port is arranged at the head end, a terminal end is connected to the first waste liquid chamber, and C sample quantitative chambers are arranged outside the channel; the first waste liquid chamber is connected to the atmosphere; The drug channel has a drug chamber at its head end, a second waste liquid chamber at its tail end, and N drug quantitative chambers at its outer side; the drug chamber is provided with a drug inlet, and the second waste liquid chamber is connected to an air hole connected to the atmosphere through an airway; M, N, and C are all positive integers, and N=M×C, and the N drug quantitative chambers correspond one to one with the M×C sample quantitative chambers; the sample quantitative chamber is arranged downstream of the corresponding drug quantitative chamber, and the top of the sample quantitative chamber is connected to the bottom of the corresponding drug quantitative chamber through a microchannel.

7. The microfluidic organoid drug screening chip according to claim 6, characterized in that: From top to bottom, it includes: cover plate, drug layer, microchannel layer, organoid layer and bottom plate; The drug channel is arranged in the drug layer; The microchannel is disposed in the microchannel layer; The injection channel is arranged in the organoid layer; The drug inlet and the sample adding port are both arranged on the cover plate; the drug inlet is matched with the head end of each drug channel, and the sample adding port is matched with the sample adding chamber; The organoid layer is also provided with an airway connected to the first waste liquid chamber, the drug layer is also provided with an airway connected to the second waste liquid chamber, and the cover plate is also provided with air holes connected to the atmosphere, and the air holes are matched with the airway ends of the drug layer and the organoid layer.

8. The microfluidic organoid drug screening chip according to claim 6 or 7, characterized in that: The volume of the sample quantitative chamber on the same injection channel gradually increases or decreases.

9. The microfluidic organoid drug screening chip according to claim 6 or 7, characterized in that: The sample quantitative chamber and the corresponding drug quantitative chamber on the same sample channel are located in the same radial direction.

10. The microfluidic organoid drug screening chip according to claim 6 or 7, characterized in that: The microchannel is in a serpentine shape.

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