A microfluidic analysis chip and analysis method based on spatial encoding

Abstract

This invention discloses a microfluidic analysis chip and analysis method based on spatial coding. The chip includes analysis units arranged around the same centrifugal axis. Each analysis unit includes: a first liquid drainage channel; the first liquid drainage channel includes one or more transverse channels connected end-to-end, with a first quantitative chamber disposed outside the transverse channels; a second liquid drainage channel; the second liquid drainage channel includes one or more longitudinal channels connected end-to-end, each longitudinal channel intersecting with each transverse channel, and is provided with N second quantitative chambers, each corresponding to a first quantitative chamber on one of the N transverse channels; and reaction chambers corresponding one-to-one with each first quantitative chamber; each first quantitative chamber and its corresponding second quantitative chamber are connected to the corresponding reaction chamber. This invention enables more complex analytical combinations without relying on fluorescent labeling, thereby accurately achieving high-throughput, multi-sample, multi-index multiplex analysis.

Description

Technical Field

[0001] This invention belongs to the field of microfluidic chips, and more specifically, relates to a microfluidic analysis chip and analysis method based on spatial coding. Background Technology

[0002] Microfluidics, as an emerging scientific technology, has been applied to many fields such as chemistry, biology, engineering, and physics. Its strong interdisciplinary nature has led to breakthroughs in the precise manipulation of time, space, and analytes, enabling the solution of many key problems in life science analysis. Microfluidics can integrate detection experiments that were previously only possible in a laboratory onto a small chip, saving not only consumable costs and time, but more importantly, integrating multiple detection technologies to improve detection efficiency.

[0003] In April 2020, Professor Pardis C. Sabeti of MIT published an article in *Nature* entitled "Massively multiplexed nucleic acid detection using Cas13," in which researchers developed a detection technology called CARMEN-Cas13, providing a solution to the current shortage of clinical testing methods. The authors first performed nucleic acid extraction, amplification, and transcription on several samples. Next, each sample was labeled with a color code using a fluorescent dye. Similarly, probes targeting several viral RNAs were prepared into detection mixtures and labeled with fluorescent dyes. Each sample and each detection mixture was prepared into an emulsion. These sample emulsions were then mixed with the emulsions of the detection solutions and loaded onto a specially designed microplate. The plate was dotted with figure-eight-shaped wells, each well just large enough to hold two microdroplets from the emulsion. The microplate was first imaged using fluorescence microscopy. Then, an electric shock was performed. The shock caused the two microdroplets in each well to fuse. If the RNA in the sample droplet can be recognized by the crRNA in the detection droplet, it will activate Cas13 in the detection droplet. This then cleaves the fluorescent probe, causing the fluorescent group to fluoresce under excitation light. Finally, a second fluorescence microscopy scan of the microplate reveals which wells emitted the probe's specific fluorescence, thus determining whether the sample contains the target nucleic acid. This method, combining microfluidic emulsification and color coding techniques with fluorescence microscopy, significantly reduces the cost of broad-spectrum pathogen detection and offers high specificity and sensitivity. However, it requires multiple fluorescent dyes and involves 1050 color codes. Samples with similar colors may be incorrectly identified as sample or test solution numbers, thus requiring a high-resolution fluorescence microscope. Furthermore, the two emulsion droplets randomly enter the "8"-shaped wells, potentially leading to fusion of identical droplets, rendering the well invalid and wasting reagents and sample.

[0004] The patent document with patent number ZL2021108258929 discloses a chip for high-throughput reagent quantitative distribution and analysis assisted by microchannels. It successfully realizes the precise injection of quantitative liquid at different radius positions by utilizing hydrophobic extension channels and effectively improves the detection throughput. However, this chip can only be used for the detection of one sample or one reagent, and still has certain limitations for the detection of multiple samples and multiple targets. Summary of the Invention

[0005] To address the shortcomings and improvement needs of existing technologies, this invention provides a microfluidic analysis chip and analysis method based on spatial coding. The purpose is to achieve more complex analysis combinations without relying on fluorescent labeling by improving the structure of the microfluidic chip, thereby accurately realizing high-throughput multi-sample and multi-index multiple analysis.

[0006] To achieve the above objectives, according to one aspect of the present invention, a microfluidic analysis chip based on spatial coding is provided, comprising: one or more analysis units arranged around the same centrifugal axis; the analysis unit includes:

[0007] At least one first liquid drainage channel; the first liquid drainage channel includes one or more transverse channels connected end to end; each first liquid drainage channel has a first sample inlet at its first end and is connected to a first waste liquid chamber at its end; a first quantitative chamber is provided outside the transverse channels, and in the same detection unit, all transverse channels are arranged sequentially from the inside to the outside.

[0008] At least one second liquid drainage channel; the second liquid drainage channel includes one or more longitudinal channels connected end to end; each second liquid drainage channel has a second sample inlet at its first end and is connected to a second waste liquid chamber at its last end; each longitudinal channel intersects with each transverse channel, and N second quantitative chambers are provided along the longitudinal channel, where N is the number of transverse channels in the same detection unit; each of the N second quantitative chambers corresponds to a first quantitative chamber on each of the N transverse channels, and the second quantitative chamber is located inside its corresponding first quantitative chamber;

[0009] And a reaction chamber that corresponds one-to-one with each of the first quantitative chambers; the first quantitative chamber and its corresponding second quantitative chamber are all connected to the corresponding reaction chamber.

[0010] In some optional embodiments, the bottom end of the first quantitative chamber is connected to its corresponding reaction chamber via a first drainage microchannel, and the bottom end of the second quantitative chamber is connected to its corresponding first quantitative chamber via a second drainage microchannel.

[0011] In some optional embodiments, the bottom end of the first quantitative chamber is connected to its corresponding reaction chamber via a first drainage microchannel, and the bottom end of the second quantitative chamber is connected to its corresponding reaction chamber via a third drainage microchannel.

[0012] In some alternative embodiments, the volume of the second metering chamber on the same second liquid drainage channel gradually increases or gradually decreases.

[0013] In some alternative embodiments, the volume of the first metering chamber on the same first liquid drainage channel gradually increases or gradually decreases.

[0014] In some optional embodiments, the analysis unit further includes: a buffer storage chamber corresponding to each of the second liquid drainage channels; the buffer storage chamber is connected to the head end of the corresponding second liquid drainage channel and stores buffer solution therein.

[0015] In some alternative embodiments, the first liquid drainage channel has a structure that expands at the end and narrows at the outlet at the inlet of the first metering chamber; this structure facilitates the extrusion of the first liquid into the first metering chamber.

[0016] According to another aspect of the present invention, an analysis method based on the above-described spatial coding-based microfluidic analysis chip is provided, comprising the following steps:

[0017] (S1) After injecting the reaction solution into the first sample inlet of each first liquid drainage channel, the microfluidic analysis chip is centrifuged at the first speed so that the reaction solution in each first liquid drainage channel enters the first quantitative chamber in sequence, and the excess reaction solution enters the first waste liquid chamber.

[0018] (S2) After injecting the reaction solution into the second sample inlet of each second liquid drainage channel, the microfluidic analysis chip is centrifuged at the second speed so that the reaction solution in each second liquid drainage channel enters the second quantitative chamber in sequence, and the excess reaction solution enters the second waste liquid chamber.

[0019] (S3) Centrifuge the microfluidic analysis chip at a third rotation speed so that the reaction liquid in each of the first and second quantitative chambers enters the corresponding reaction chamber and mixes and reacts.

[0020] (S4) After the reaction is completed, the signals in each reaction chamber are read, and the reaction results under different reaction liquid combinations are analyzed based on the correspondence between the position information of the reaction chambers and the reaction liquid combinations.

[0021] The reaction solutions in the first and second quantitative chambers corresponding to the same reaction chamber are different types of reaction solutions; the location information of the reaction chamber includes the transverse channel number of the first quantitative chamber and the longitudinal channel number of the second quantitative chamber; the first rotation speed and the second rotation speed are both less than the third rotation speed.

[0022] In some optional embodiments, if the detection unit in the microfluidic chip does not include a buffer storage chamber, then the step between step (S2) and step (S3) further includes:

[0023] After injecting a preset volume of buffer solution into the second sample inlet of each second liquid drainage channel, the microfluidic chip is centrifuged at the fourth rotation speed so that the buffer solution in each second liquid drainage channel comes into contact with the reaction liquid in the second quantitative chamber in sequence along the second liquid drainage channel and finally flows into the second waste liquid chamber.

[0024] The fourth rotational speed is less than the third rotational speed.

[0025] In some optional embodiments, if the detection unit in the microfluidic chip includes a buffer storage chamber, then the following step is further included between step (S2) and step (S3):

[0026] The buffer solution in each buffer storage chamber is released into the corresponding second liquid drainage channel. Then, the microfluidic chip is centrifuged at the fifth rotation speed so that the buffer solution in each second liquid drainage channel comes into contact with the reaction liquid in the second quantitative chamber in sequence along the second liquid drainage channel and finally flows into the second waste liquid chamber.

[0027] The fifth rotational speed is less than the third rotational speed.

[0028] In summary, the above-described technical solutions conceived in this invention can achieve the following beneficial effects:

[0029] The microfluidic analysis chip based on spatial coding provided by this invention includes a liquid drainage channel arranged laterally and longitudinally in a cross-combination layout for each analysis unit. Simultaneously, corresponding quantitative chambers are provided on the lateral and longitudinal channels, and two corresponding quantitative chambers are connected to the same reaction chamber. Through this channel layout, this invention can control the combination of reaction solutions in both lateral and longitudinal dimensions using centrifugation, thereby enabling more complex analytical combinations. Specifically, this invention can mix the same sample with different reagents, the same reagent with different samples, or the same reagent at different concentrations, thereby increasing analytical throughput and efficiency. Furthermore, the cross-combination channel design of this invention ensures that each reaction chamber has a positional coding information corresponding to a reaction solution combination, allowing the determination of the corresponding reaction solution combination based on the positional information of the reaction chamber. This enables accurate high-throughput, multi-sample, multi-index multiplex analysis without relying on fluorescent labeling. Attached Figure Description

[0030] Figure 1 This is a schematic diagram of a microfluidic analysis chip based on spatial coding provided in Embodiment 1 of the present invention;

[0031] Figure 2 This is a schematic diagram of the analysis unit in the microfluidic analysis chip based on spatial coding provided in Embodiment 1 of the present invention;

[0032] Figure 3 This is a schematic diagram of the microfluidic analysis chip based on spatial coding provided in Embodiment 1 of the present invention during the actual analysis process; wherein, (a) to (c) are different states of the analysis unit during the analysis process;

[0033] Figure 4 This is a schematic diagram of the microfluidic analysis chip based on spatial coding provided in Embodiment 1 of the present invention during another actual analysis process; wherein, (a) to (d) are different states of the analysis unit during the analysis process;

[0034] Figure 5 This is a schematic diagram of each chip layer in the microfluidic analysis chip based on spatial coding provided in Embodiment 1 of the present invention; wherein, (a) is the top cover layer, (b) is the connecting layer, (c) is the longitudinal channel layer, (d) is the transverse channel layer, and (e) is the bottom cover layer;

[0035] Figure 6 This is a schematic diagram of the analysis method provided in Embodiment 2 of the present invention; wherein (a) to (d) represent different states of the analysis unit during the analysis process;

[0036] Figure 7 This is a schematic diagram of the analysis method provided in Embodiment 3 of the present invention; wherein (a) to (h) represent different states during the contact process between the buffer solution and the liquid in the second quantitative chamber;

[0037] Figure 8 This is a schematic diagram of the analysis unit in the microfluidic analysis chip based on spatial coding provided in Embodiment 4 of the present invention;

[0038] Figure 9 This is a schematic diagram of the analysis unit in the microfluidic analysis chip based on spatial coding provided in Embodiment 7 of the present invention;

[0039] Figure 10 This is a schematic diagram of a microfluidic analysis chip based on spatial coding provided in Embodiment 8 of the present invention;

[0040] Figure 11 This is a partial schematic diagram of the first drainage channel in the microfluidic analysis chip based on spatial coding provided in Embodiment 8 of the present invention;

[0041] In all the accompanying drawings, the same reference numerals are used to denote the same elements or structures, wherein:

[0042] 1-Centrifugal shaft;

[0043] 2-First liquid drainage channel; 21-Transverse flow channel; 22-First sample inlet; 23-First waste liquid chamber; 24-First quantitative chamber;

[0044] 3-Second liquid drainage channel; 31-Longitudinal flow channel; 32-Second sample inlet; 33-Second waste liquid chamber; 34-Second quantitative chamber;

[0045] 4-Reaction chamber;

[0046] 5-First drainage microchannel;

[0047] 6-Second drainage microchannel;

[0048] 7-Third drainage microchannel. Detailed Implementation

[0049] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention. Furthermore, the technical features involved in the various embodiments of this invention described below can be combined with each other as long as they do not conflict with each other.

[0050] In this invention, the terms "first," "second," etc. (if present), used in the invention and accompanying drawings are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. Furthermore, in the description of this invention, terms such as "upper," "lower," "left," "right," "inner," and "outer," indicating directional or positional relationships, are based on the directional or positional relationships shown in the accompanying drawings. This is merely for ease of description and does not indicate or imply that the device or element must have a specific orientation, or be constructed and operated in a specific orientation; therefore, it should not be construed as a limitation of the invention. Moreover, it should be noted that in the description of this invention, unless otherwise explicitly specified and limited, the terms "installed," "connected," and "linked" should be interpreted broadly. For example, they can refer to a fixed connection, a detachable connection, or an integral connection; they can refer to a mechanical connection or an electrical connection; they can refer to a direct connection or an indirect connection through an intermediate medium; and they can refer to the internal connection of two elements. Those skilled in the art can understand the specific meaning of the above terms in this invention according to the specific circumstances.

[0051] It is easy to understand that in centrifugal chips, terms such as "upstream," "downstream," "high," and "low" are all relative to the distance from the centrifugal axis. Specifically, points closer to the centrifugal axis are located "upstream," while points farther from the centrifugal axis are located "downstream." Points closer to the centrifugal axis are higher, while points farther from the centrifugal axis are lower.

[0052] To achieve more complex analytical combinations without relying on fluorescent labeling, this invention provides a microfluidic analysis chip and method based on spatial coding. The overall idea is to improve the structure of the centrifugal microfluidic chip so that the channels for guiding the reaction liquid are arranged in a cross-combination layout, thereby achieving more complex analytical combinations. At the same time, under the cross-combination layout, each reaction chamber has position coding information that corresponds one-to-one with the reaction liquid combination, so as to accurately achieve high-throughput multi-sample, multi-index multiple analysis without relying on fluorescent labeling.

[0053] The following is an example.

[0054] Example 1:

[0055] A microfluidic analysis chip based on spatial coding, such as Figure 1 As shown, it includes: one or more analysis units arranged around the same centrifugal axis 1; as Figure 2 As shown, the analysis unit includes:

[0056] Three first liquid drainage channels 2; each first liquid drainage channel 2 includes a transverse flow channel 21; each first liquid drainage channel 2 has a first sample inlet 22 at its head and is connected to a first waste liquid chamber 23 at its end; a first quantitative chamber 24 is provided outside the transverse flow channel 21, and in the same detection unit, all transverse flow channels 21 are arranged sequentially from the inside to the outside.

[0057] Four second liquid drainage channels 3; each second liquid drainage channel 3 includes a longitudinal flow channel 31; each second liquid drainage channel 3 has a second sample inlet 32 ​​at its first end and is connected to a second waste liquid chamber 33 at its last end; each longitudinal flow channel 31 intersects with each transverse flow channel within the same analytical unit, and three second quantitative chambers 34 are provided along the longitudinal flow channel 31; each of the three second quantitative chambers 34 corresponds to a first quantitative chamber on one of the three transverse flow channels 21, and the second quantitative chamber 34 is located inside its corresponding first quantitative chamber 24; for example Figure 2 As shown, in order to facilitate the entry of liquid into the second metering chamber 34, in a preferred embodiment, each longitudinal channel 31 extends from the proximal end to the distal end in a "z" shape.

[0058] And a reaction chamber 4 is provided in a one-to-one correspondence with each of the first quantitative chambers 24; the bottom end of the first quantitative chamber 24 is connected to its corresponding reaction chamber 4 through the first drainage microchannel 5, and the bottom end of the second quantitative chamber 34 is connected to its corresponding first quantitative chamber 24 through the second drainage microchannel 6.

[0059] It should be noted that the number of first liquid drainage channels, the number of transverse channels included in the first liquid drainage channel, the number of second liquid drainage channels, and the number of longitudinal channels included in the second liquid drainage channel in this embodiment are all illustrative examples and should not be construed as the sole limitation of the present invention. In other embodiments of the present invention, the number of first liquid drainage channels in the analysis unit may also be set to other numbers, or even only one first liquid drainage channel may be set. Similarly, the number of second liquid drainage channels in the analysis unit may also be set to other numbers, or even only one second liquid drainage channel may be set. The first liquid drainage channel may also be configured to include multiple transverse channels connected end to end, and similarly, the second liquid drainage channel may also be configured to include multiple longitudinal channels connected end to end.

[0060] The microfluidic analysis chip based on spatial coding provided in this embodiment can complete the spatial position coding of each reaction chamber according to the number of the transverse channel where the first quantitative chamber connected to the reaction chamber is located and the number of the longitudinal flow channel where the second quantitative chamber connected to the reaction chamber is located. Each reaction chamber corresponds to a reaction combination, including the reaction liquid involved in the combination and the corresponding concentration.

[0061] In this embodiment, the first quantitative chambers on each first liquid drainage channel have the same volume, and the second quantitative chambers on each second liquid drainage channel also have the same volume. In other embodiments of the present invention, the volume of the second quantitative chambers on the same second liquid drainage channel may be gradually increased or gradually decreased, and the volume of the first quantitative chambers on the same first liquid drainage channel may be gradually increased or gradually decreased, thereby obtaining different concentration gradients.

[0062] In this embodiment, there are a total of 3 transverse flow channels and 4 longitudinal flow channels, which can form 12 combinations. The transverse flow channels are used as sample flow channels, and the longitudinal flow channels are used as drug flow channels. H1, H2, and H3 represent samples introduced into the 3 transverse flow channels, and V1, V2, V3, and V4 represent drugs introduced into the 4 longitudinal flow channels. The 12 combinations can be represented as follows:

[0063] H1-V1, H1-V2, H1-V3, H1-V4, H2-V1, H2-V2, H2-V3, H2-V4, H3-V1, H3-V2, H3-V3, H3-V4.

[0064] Taking H1-V1 as an example, it can be understood as detecting the V1 index of sample H1. During low-speed centrifugation, sample H1 can be sequentially introduced into the first quantitative chamber along the first liquid drainage channel to complete sample quantification. Similarly, drug V1 can be sequentially introduced into the second quantitative chamber along the second liquid drainage channel to complete drug quantification. After high-speed centrifugation, since sample H1 is located outside the first quantitative chamber where the corresponding V1 is located, it experiences a greater centrifugal force and is first transferred to the downstream reaction chamber. At this time, drug V1 enters the first quantitative chamber where H1 is located through the hydrophobic drainage microvalve and then transfers to the downstream reaction chamber.

[0065] Taking the introduction of sample solution through the first liquid inlet channel and drug solution through the second liquid drainage channel as an example, the microfluidic analysis chip based on spatial coding provided in this embodiment can be operated as follows during actual analysis: First, the sample solution is introduced through the first liquid inlet channel, and the chip is centrifuged at low speed to allow the sample solution to enter the first quantitative chamber for sample quantification. Figure 3 As shown in (a) in the text;

[0066] Then, the drug solution is introduced through the second liquid drainage channel, and the chip is centrifuged at low speed to allow the drug solution to enter the second quantitative chamber for drug quantification. Figure 3 As shown in (b) in the image;

[0067] Finally, the chip is centrifuged at high speed to transfer the sample in the first quantitative chamber and the drug in the second quantitative chamber to the downstream reaction chamber, thus completing the reaction.

[0068] Similarly, when the sample solution is introduced into the first liquid inlet channel and the drug solution is introduced into the second liquid drainage channel, the microfluidic analysis chip based on spatial coding provided in this embodiment can also be operated according to the following steps when actually used for analysis:

[0069] First, the sample solution is introduced through the first liquid inlet channel, and the chip is centrifuged at low speed to allow the sample solution to enter the first quantitative chamber for sample quantification. Figure 4 As shown in (a) in the text;

[0070] Then, the chip is centrifuged at high speed to transfer the sample solution in the first quantitative chamber to the downstream reaction chamber, such as... Figure 4 As shown in (b) in the image;

[0071] Next, the drug solution is introduced through the second liquid drainage channel, and the chip is centrifuged at low speed to allow the drug solution to enter the second quantitative chamber for drug quantification. Figure 4 As shown in (c);

[0072] Finally, the chip is centrifuged at high speed to transfer the drug solution in the second quantitative chamber to the downstream reaction chamber, where it reacts with the sample. Figure 4 As shown in (d) in the figure.

[0073] Optionally, such as Figure 5 As shown, the microfluidic analysis chip based on spatial coding provided in this embodiment has a five-layer structure, from top to bottom: top cap layer, connecting layer, longitudinal channel layer, transverse channel layer, and bottom cap layer, as shown in the figures below. Figure 5 As shown in (a), (b), (c), (d), and (e);

[0074] Both the first and second drainage microchannels are located in the connecting layer;

[0075] The second liquid drainage channel and the second metering chamber thereon, as well as the second waste liquid chamber, are all located in the longitudinal flow channel layer;

[0076] The first liquid drainage channel and the first metering chamber thereon, as well as the first waste liquid chamber, are all located in the transverse flow channel layer;

[0077] The first sample inlet is set through the top cover layer, the connecting layer, and the longitudinal flow channel layer; the second sample inlet is set through the top cover layer and the connecting layer.

[0078] It should be noted that the layered design of the chip in this embodiment is only one optional implementation. In other embodiments of the present invention, other layered design schemes may also be adopted while ensuring the communication relationship between the chambers.

[0079] In summary, the spatially encoded microfluidic analysis chip provided in this embodiment includes horizontally and vertically arranged liquid drainage channels in each analysis unit, forming a cross-combination layout. Simultaneously, corresponding quantitative chambers are provided on the horizontal and vertical channels, and two corresponding quantitative chambers are connected to the same reaction chamber. Through this channel layout, this embodiment can control the combination of reaction solutions in both horizontal and vertical dimensions using centrifugation, thereby enabling more complex analytical combinations. Specifically, this embodiment can mix the same sample with different reagents, the same reagent with different samples, or the same reagent at different concentrations, thereby increasing analytical throughput and efficiency. Furthermore, the cross-combination channel design in this embodiment ensures that each reaction chamber has a positional encoding information corresponding to a reaction solution combination, allowing the determination of the corresponding reaction solution combination based on the positional information of the reaction chamber. This enables high-throughput, multi-sample, multi-index multiplex analysis without relying on fluorescent labeling.

[0080] This embodiment implements spatial encoding technology and combines it with microchannel assistance. It utilizes fixed-volume quantitative chambers to quantify different samples (M samples) and detection reagents (N reagents) separately. Then, using centrifugation, different combinations of samples and detection reagents are fixed in the detection wells, eliminating crosstalk between wells and improving specificity. The spatial encoding technology used in this embodiment physically separates the M×N reaction chambers, thus eliminating the need for multiple fluorescent signal probes. Only one signal reporter probe is needed to determine the presence of the target analyte in the sample, greatly simplifying the signal readout method and reducing detection costs. The microfluidic analysis chip provided in this embodiment, utilizing spatial encoding technology, enables the simultaneous detection of multiple samples and multiple targets within a single chip, significantly reducing sample detection waiting time, especially in resource-limited areas. This further contributes to promoting the application of microfluidic chips in POCT. Furthermore, this embodiment has wide applications, including drug screening, bacterial drug sensitivity testing, organoid culture, nucleic acid detection, and immunoassay, making it a POCT detection tool with enormous development potential.

[0081] Example 2:

[0082] Based on the analysis method of the microfluidic analysis chip based on spatial coding provided in Embodiment 1 above, such as Figure 6 As shown, it includes the following steps:

[0083] (S1) After injecting the reaction solution into the first sample inlet 22 of each first liquid drainage channel 2, the microfluidic analysis chip is centrifuged at a first rotation speed so that the reaction solution in each first liquid drainage channel 2 sequentially enters the first quantitative chamber 24, and the excess reaction solution enters the first waste liquid chamber 23; such as Figure 6 As shown in (a) in the text;

[0084] Optionally, in this embodiment, the reaction solution injected into the three first liquid drainage channels 2 is three different types of sample solutions; the first rotation speed can allow the liquid to enter the first quantitative chamber, but will not break through the first drainage microchannel;

[0085] (S2) After injecting the reaction solution into the second sample inlet 32 ​​of each of the second liquid drainage channels 3, the microfluidic analysis chip is centrifuged at the second rotation speed so that the reaction solution in each of the second liquid drainage channels 3 sequentially enters the second quantitative chamber 34, and the excess reaction solution enters the second waste liquid chamber 33; such as Figure 6 As shown in (b) in the image;

[0086] Optionally, in this embodiment, the reaction liquid injected into the four second liquid drainage channels 3 is four different kinds of drug solutions; the second rotation speed can allow the liquid to enter the second metering chamber, but will not break through the second drainage microchannel;

[0087] (S3) Centrifuge the microfluidic analysis chip at a third rotation speed so that the reaction liquid in each of the first quantitative chamber 24 and the second quantitative chamber 34 enters the corresponding reaction chamber 4 to mix and react.

[0088] During centrifugation, the centrifugal force on the first metering chamber 24 is greater than that on the corresponding second metering chamber 34. Therefore, the reaction solution in the first metering chamber 24 will first enter the downstream reaction chamber 4, and then the reaction solution in the corresponding second metering chamber 34 will enter the corresponding first metering chamber 24, and then enter the downstream reaction chamber 4, as follows. Figure 6 As shown in (c) and (d) in the text;

[0089] The third rotation speed is greater than the first and second rotation speeds, and it allows the liquid in the first and second metering chambers to break through the drainage microchannels and enter the reaction chamber;

[0090] (S4) After the reaction is completed, the signals in each reaction chamber 4 are read, and the reaction results under different reaction liquid combinations are analyzed based on the correspondence between the position information of the reaction chambers and the reaction liquid combinations.

[0091] Different combinations of reaction solutions yield different reaction results. By reading these results as signals, the reaction status corresponding to the specific combination of reaction solutions can be determined.

[0092] This embodiment can analyze the reaction between different reaction solutions.

[0093] In practical applications, the specific method of introducing samples or solutions can be set according to actual analytical needs, as long as the reaction solutions in the first and second quantitative chambers corresponding to the same reaction chamber are different types of reaction solutions. In other embodiments of the present invention, the same or different types of sample solutions can be introduced through the first liquid drainage channel, while different concentrations of the same drug can be introduced through the second liquid drainage channel, thereby allowing analysis of the effect of drug concentration on the reaction; in other embodiments of the present invention, the same or different types of drug solutions can be introduced through the second liquid drainage channel, while different concentrations of the same sample solution can be introduced through the first liquid drainage channel, thereby allowing analysis of the effect of sample concentration on the reaction;

[0094] In other embodiments of the present invention, the drug solution may be introduced through the first liquid drainage channel and the sample solution may be introduced through the second liquid drainage channel.

[0095] Example 3:

[0096] The analysis method for the microfluidic analysis chip based on spatial coding provided in Embodiment 1 above. This embodiment is similar to Embodiment 2 above, except that in this embodiment, the method further includes the following step between step (S2) and step (S3):

[0097] After injecting a preset volume of buffer solution into the second sample inlet of each second liquid drainage channel, the microfluidic chip is centrifuged at the fourth rotation speed so that the buffer solution in each second liquid drainage channel comes into contact with the reaction liquid in the second quantitative chamber in sequence along the second liquid drainage channel and finally flows into the second waste liquid chamber.

[0098] The fourth rotation speed is less than the third rotation speed, which allows the buffer solution to flow along the longitudinal flow channel and come into contact with the reaction liquid in the second quantitative chamber in sequence, without allowing the liquid in the first and second quantitative chambers to enter the reaction chamber.

[0099] It is easy to understand that the buffer solution is a liquid that does not affect the properties of the reagent. Optionally, in this embodiment, the buffer solution used is an isotonic solution of the sample to be tested.

[0100] like Figure 7As shown in (a) to (h), during the contact of the buffer solution with each second quantitative chamber, a certain amount of solution exchange occurs, resulting in a decrease in the liquid concentration in the second quantitative chamber and an increase in the drug concentration in the buffer solution. As the buffer solution flows downstream in sequence, the drug concentration gradient in the second quantitative chamber increases sequentially from the inside to the outside, thereby realizing the online formation of the drug concentration gradient.

[0101] Example 4:

[0102] A microfluidic analysis chip based on spatial coding. This embodiment is similar to Embodiment 1 above, except that, as Figure 8 As shown, in this embodiment, the bottom end of the first quantitative chamber 24 is connected to its corresponding reaction chamber 4 through the first drainage microchannel 5, and the bottom end of the second quantitative chamber 34 is directly connected to its corresponding reaction chamber 4 through the third drainage microchannel 7.

[0103] The method used for analysis and detection in this embodiment is the same as that in Embodiment 1 above. For details, please refer to the descriptions of Embodiments 1 to 3 above.

[0104] Example 5:

[0105] A microfluidic analysis chip based on spatial coding. This embodiment is similar to Embodiment 1 above, except that in this embodiment, the analysis unit further includes: a buffer storage chamber corresponding to each of the second liquid drainage channels; the buffer storage chamber is connected to the head end of the corresponding second liquid drainage channel and stores buffer solution therein.

[0106] This embodiment features a separate buffer storage chamber corresponding to the second liquid drainage channel, which can store a predetermined volume of buffer solution in advance, facilitating the online formation of drug concentration gradients.

[0107] Optionally, the buffer storage chamber is connected to the beginning of the corresponding second liquid drainage channel via a microvalve structure such as a sacrificial or dissolvable microvalve.

[0108] Example 6:

[0109] The analysis method for the microfluidic analysis chip based on spatial coding provided in Embodiment 5 above. This embodiment is similar to Embodiment 2 above, except that in this embodiment, the method further includes the following step between step (S2) and step (S3):

[0110] The buffer solution in each buffer storage chamber is released into the corresponding second liquid drainage channel. Then, the microfluidic chip is centrifuged at the fifth rotation speed so that the buffer solution in each second liquid drainage channel comes into contact with the reaction liquid in the second quantitative chamber in sequence along the second liquid drainage channel and finally flows into the second waste liquid chamber.

[0111] The fifth rotation speed is less than the third rotation speed, which allows the buffer solution to flow along the longitudinal flow channel and come into contact with the reaction liquid in the second quantitative chamber in sequence, without allowing the liquid in the first and second quantitative chambers to enter the reaction chamber.

[0112] Example 7:

[0113] A microfluidic analysis chip based on spatial coding. This embodiment is similar to Embodiment 1 above, except that, as Figure 9 As shown, in this embodiment, the number of first liquid drainage channels in the analysis unit is 1, and the first liquid drainage channel includes 3 transverse flow channels connected end to end. The entire first liquid drainage channel is "S" shaped and extends from the proximal end to the distal end.

[0114] Example 8:

[0115] A microfluidic analysis chip based on spatial coding. This embodiment is similar to Embodiment 1 above, except that, in this embodiment, as... Figure 10 and Figure 11 As shown, the first liquid drainage channel has a structure that expands at the end and narrows at the outlet at the inlet of the first metering chamber, which makes it easy for the first liquid to be squeezed into the first metering chamber.

[0116] The method used for analysis and detection in this embodiment is the same as that in Embodiment 1 above. For details, please refer to the descriptions of Embodiments 1 to 3 above.

[0117] Those skilled in the art will readily understand that the above description is merely 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 within the scope of protection of the present invention.

Claims

1. A microfluidic analysis chip based on spatial coding, characterized in that, include: One or more analysis units arranged around the same centrifugal axis; the analysis unit includes: At least one primary fluid drainage channel; The first liquid drainage channel includes a transverse channel or a channel formed by connecting multiple transverse channels end to end, extending from the proximal end in an "S" shape to the distal end; each first liquid drainage channel has a first sample inlet at its beginning and is connected to a first waste liquid chamber at its end; a first quantitative chamber is provided outside the transverse channel, and in the same detection unit, all transverse channels are arranged sequentially from the inside to the outside; At least one second liquid drainage channel; the second liquid drainage channel includes a longitudinal flow channel extending from the proximal end to the distal end in a "Z" shape; each second liquid drainage channel has a second sample inlet at its first end and is connected to a second waste liquid chamber at its last end; the longitudinal flow channel intersects with each transverse flow channel respectively, and is provided along the longitudinal flow channel. N A second quantitative chamber, N The number of transverse flow channels in the same detection unit; N Each of the second quantitative chambers is respectively connected to N A first metering chamber corresponds to a second metering chamber located inside the corresponding first metering chamber on each transverse flow channel; the volume of the second metering chamber on the same second liquid drainage channel gradually increases or gradually decreases. And a reaction chamber that corresponds one-to-one with each of the first quantitative chambers; the first quantitative chamber and its corresponding second quantitative chamber are all connected to the corresponding reaction chamber.

2. The microfluidic analysis chip based on spatial coding as described in claim 1, characterized in that, The bottom end of the first quantitative chamber is connected to its corresponding reaction chamber through a first drainage microchannel, and the bottom end of the second quantitative chamber is connected to its corresponding first quantitative chamber through a second drainage microchannel.

3. The microfluidic analysis chip based on spatial coding as described in claim 1, characterized in that, The bottom end of the first quantitative chamber is connected to its corresponding reaction chamber through a first drainage microchannel, and the bottom end of the second quantitative chamber is connected to its corresponding reaction chamber through a third drainage microchannel.

4. The microfluidic analysis chip based on spatial coding as described in any one of claims 1 to 3, characterized in that, The volume of the first quantitative chamber on the same first liquid drainage channel gradually increases or gradually decreases.

5. The microfluidic analysis chip based on spatial coding as described in any one of claims 1 to 3, characterized in that, The analysis unit further includes a buffer storage chamber corresponding to each of the second liquid drainage channels; the buffer storage chamber is connected to the head end of the corresponding second liquid drainage channel and stores buffer solution therein.

6. The microfluidic analysis chip based on spatial coding as described in any one of claims 1 to 3, characterized in that, The first liquid drainage channel has a structure that expands at the end and narrows at the outlet at the inlet of the first quantitative chamber.

7. An analysis method based on the microfluidic analysis chip based on spatial coding as described in any one of claims 1 to 6, characterized in that, Includes the following steps: (S1) After injecting the reaction solution into the first sample inlet of each first liquid drainage channel, the microfluidic analysis chip is centrifuged at a first speed so that the reaction solution in each first liquid drainage channel enters the first quantitative chamber in sequence, and the excess reaction solution enters the first waste liquid chamber. (S2) After injecting the reaction solution into the second sample inlet of each second liquid drainage channel, the microfluidic analysis chip is centrifuged at the second speed so that the reaction solution in each second liquid drainage channel enters the second quantitative chamber in sequence, and the excess reaction solution enters the second waste liquid chamber. (S3) The microfluidic analysis chip is centrifuged at a third rotation speed so that the reaction liquid in each of the first and second quantitative chambers enters the corresponding reaction chamber for mixing and reaction; (S4) After the reaction is completed, the signals in each reaction chamber are read, and the reaction results under different reaction liquid combinations are analyzed based on the correspondence between the position information of the reaction chambers and the reaction liquid combinations. The reaction solutions in the first and second quantitative chambers corresponding to the same reaction chamber are different types of reaction solutions; the location information of the reaction chamber includes the transverse channel number of the first quantitative chamber and the longitudinal channel number of the second quantitative chamber; the first rotation speed and the second rotation speed are both less than the third rotation speed.

8. The analytical method as described in claim 7, characterized in that, If the detection unit in the microfluidic analysis chip does not include a buffer storage chamber, then the step between step (S2) and step (S3) further includes: After injecting a preset volume of buffer solution into the second sample inlet of each second liquid drainage channel, the microfluidic analysis chip is centrifuged at a fourth rotation speed so that the buffer solution in each second liquid drainage channel comes into contact with the reaction liquid in the second quantitative chamber in sequence along the second liquid drainage channel and finally flows into the second waste liquid chamber. The fourth rotational speed is less than the third rotational speed.

9. The analytical method as described in claim 7, characterized in that, If the detection unit in the microfluidic analysis chip includes a buffer storage chamber, then the step between step (S2) and step (S3) further includes: The buffer solution in each buffer storage chamber is released into the corresponding second liquid drainage channel. Then, the microfluidic analysis chip is centrifuged at a fifth rotation speed so that the buffer solution in each second liquid drainage channel comes into contact with the reaction liquid in the second quantitative chamber in sequence along the second liquid drainage channel and finally flows into the second waste liquid chamber. The fifth rotational speed is less than the third rotational speed.

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