A centrifugal microfluidic high-throughput analysis chip

Abstract

The application discloses a centrifugal micro-fluidic high-throughput analysis chip, and belongs to the technical field of micro-fluidic chips, which is provided with a rotating center, and comprises: M detection units arranged around the rotating center; the detection unit comprises: a waste liquid chamber; N linear quantitative distribution channels which extend in the radial direction and are provided with a first sample adding port at the leading end and connected with the waste liquid chamber at the tail end; and K reaction chambers which are distributed in the radial direction from inside to outside; the linear quantitative distribution channel is provided with K first quantitative chambers which are connected with the K reaction chambers in sequence in the direction from the rotating center to the edge through hydrophobic micro-channels; the first quantitative chamber and the reaction chamber which are connected with each other are characterized in that the reaction chamber is farther away from the rotating center. The distribution of the chambers in the detection unit of the micro-fluidic chip and the connection mode between the chambers are improved, so that the resistance and centrifugal force of the quantitative chamber are matched during centrifugation, liquid quantification is accurately realized, and chip processing is facilitated.

Description

A centrifugal microfluidic high-throughput analysis chip Technical Field

[0001] This invention belongs to the field of microfluidic chip technology, and more specifically, relates to a centrifugal microfluidic high-throughput analysis chip. Background Technology

[0002] Microfluidics, as an emerging scientific technology, is highly integrated, effectively saving on consumable and time costs and greatly improving detection efficiency. Currently, this technology is widely used in 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 it to solve many key problems in life analysis.

[0003] However, the current microfluidic chip manufacturing process is complex and has a low yield rate, directly affecting the stability of chip detection. Furthermore, the complex quantitative dispensing process requires specialized personnel, making it difficult to use. In addition, existing microfluidic chips suffer from poor integration, high reagent consumption, slow detection speed, and high detection costs.

[0004] To address the aforementioned issues, existing solutions propose setting up quantitative chambers of specific volumes along the liquid distribution channels. These quantitative chambers are connected to the reaction chambers via hydrophobic microchannels. Through the combined effect of centrifugal force and the valve-controlled action of the hydrophobic microchannels, precise liquid quantification is achieved. For example, Chinese invention patent application number 2021108258929 discloses a chip for microchannel-assisted high-throughput reagent quantitative distribution and analysis. The chip's detection unit includes reagent quantitative distribution channels arranged from the inside out relative to the rotation center; several quantitative chambers are arranged on the outer wall of the liquid distribution channel for quantitative distribution; two adjacent reagent quantitative distribution channels are connected via hydrophobic microchannels. The hydrophobic microchannels can generate resistance to liquid reagents or samples. By controlling the rotation speed to regulate the centrifugal force and the relationship between the centrifugal force and resistance, the conductivity of the hydrophobic microchannels to the reagents or samples can be controlled, thus achieving both quantitative distribution and mixing of the quantitatively distributed reagents or samples. This approach enables high-throughput quantitative dispensing and mixing of reagents or samples by setting up hydrophobic microchannels, eliminating the need to integrate a large number of valves on the chip.

[0005] In the aforementioned microfluidic chip, the dispensing channels are arranged along the circumference, and correspondingly, the quantitative chambers with the same dispensing channels are located on the same circumference. To achieve mixing of multiple reagents, multiple dispensing channels distributed along different circumferences need to be set in the same detection unit, and corresponding quantitative chambers are also distributed along different circumferences. However, as the distance from the center of rotation increases, the centrifugal force on the solution increases, and the requirements for valve control become more stringent. It is necessary to manually adjust the length of the microchannels that provide valve control, and even to make the microchannels bend in shape, which is not conducive to manufacturing and may also affect the ability of the microchannels to perform their valve control function. Summary of the Invention

[0006] To address the shortcomings and improvement needs of existing technologies, this invention provides a centrifugal microfluidic high-throughput analysis chip. The purpose is to improve the distribution of the internal chambers of the detection unit in the microfluidic chip and the connection method between the chambers, so that during centrifugation, the microchannels connected to the quantitative chamber provide resistance that matches the centrifugal force, thereby achieving accurate liquid quantification while facilitating chip fabrication.

[0007] To achieve the above objectives, the present invention provides a centrifugal microfluidic high-throughput analysis chip, which has a rotation center and includes M detection units arranged around the rotation center.

[0008] The detection unit includes: a waste liquid chamber, N linear quantitative distribution channels, and K reaction chambers;

[0009] K reaction chambers are distributed radially from the inside to the outside;

[0010] The linear quantitative distribution channel extends radially, with its inner and outer ends forming the beginning and end, respectively. The beginning end is provided with a first sample inlet, and the end is connected to the waste liquid chamber. K first quantitative chambers are provided on the linear quantitative distribution channel. The K first quantitative chambers are connected to K reaction chambers in sequence from the rotation center to the edge through hydrophobic microchannels.

[0011] In the interconnected first metering chamber and reaction chamber, the distance from the reaction chamber to the rotation center is greater than the distance from the first metering chamber to the rotation center;

[0012] Where M, N and K are all positive integers, and K>1.

[0013] In some optional embodiments, each reaction chamber in the detection unit is further provided with a quantitative sample addition unit on the side near the center of rotation; the quantitative sample addition unit includes: an independent sample addition port, and an independent quantitative chamber and an independent waste liquid chamber connected to the independent sample addition port; each independent quantitative chamber is connected to the corresponding reaction chamber through a capillary microchannel.

[0014] In some optional embodiments, the detection unit further includes: a zigzag quantitative dispensing channel extending from the rotation center to the edge;

[0015] The inner and outer ends of the zigzag quantitative distribution channel constitute its head and tail ends, respectively. The head end is provided with a second sample inlet, and the tail end is connected to the waste liquid chamber. The zigzag quantitative distribution channel is provided with K second quantitative chambers. The K second quantitative chambers are connected to the K reaction chambers in sequence along the direction from the rotation center to the edge through hydrophobic microchannels.

[0016] In the interconnected second quantitative chamber and reaction chamber, the distance from the reaction chamber to the rotation center is greater than the distance from the second quantitative chamber to the rotation center.

[0017] Furthermore, the centrifugal microfluidic high-throughput analysis chip provided by the present invention further includes: a storage chamber disposed between the rotation center and the M detection units, wherein a common sample dispensing port is provided thereon, and the second sample dispensing port of the zigzag quantitative dispensing channel in the detection unit is connected to the storage chamber.

[0018] Furthermore, the connection between the storage chamber and the second sample application port is designed as an outward protrusion.

[0019] In some optional embodiments, the detection unit further includes K sealed gas chambers corresponding one-to-one with the K reaction chambers, and the end of each reaction chamber away from the center of rotation is connected to the corresponding sealed gas chamber through a hydrophobic microchannel.

[0020] Furthermore, the hydrophobic microchannel connecting the sealed gas chamber and the reaction chamber is zigzag-shaped.

[0021] In some optional implementations, the detection unit further includes K temporary liquid storage chambers corresponding one-to-one with the K reaction chambers, and the end of each reaction chamber away from the center of rotation is connected to the corresponding temporary liquid storage chamber through a capillary channel.

[0022] In some alternative embodiments, the central region of the reaction chamber is hydrophilic, while the region outside the central region is hydrophobic.

[0023] Furthermore, in each linear quantitative dispensing channel, the quantitative chamber closest to the head is the sacrificial chamber.

[0024] In summary, the above-described technical solutions conceived in this invention can achieve the following beneficial effects:

[0025] (1) The centrifugal microfluidic high-throughput analysis chip provided by the present invention comprises multiple reaction chambers radially distributed in each unit, a quantitative dispensing channel radially distributed, and multiple quantitative chambers also radially distributed thereon. These quantitative chambers and reaction chambers are sequentially connected along the rotation center towards the edge via hydrophobic microchannels. This design increases the distance between the quantitative chambers farther from the rotation center and the connected reaction chambers, resulting in a longer hydrophobic microchannel distance between them, providing greater resistance during centrifugation. Through this structural design, the microchannel resistance experienced by the quantitative chambers matches the centrifugal force during chip operation, effectively improving valve control stability. Furthermore, the microchannel structure requires no special design, facilitating chip fabrication.

[0026] (2) In the preferred embodiment of the centrifugal microfluidic high-throughput analysis chip provided by the present invention, each reaction chamber is provided with a quantitative sample addition unit on the side near the center of rotation. This allows each reaction chamber to be injected independently, and the independent quantitative chamber in the quantitative sample addition unit can achieve accurate quantification. Compared with chips that can only pre-embed reaction reagents in the reaction chamber, this structural design in the present invention allows for flexible and accurate mixed reactions in different reaction chambers within the same detection unit, meeting various detection needs.

[0027] (3) In a preferred embodiment of the centrifugal microfluidic high-throughput analysis chip provided by the present invention, the detection unit is further provided with a zigzag quantitative distribution channel extending from the rotation center to the edge, and a plurality of quantitative chambers are provided thereon, which are connected one-to-one with a plurality of reaction chambers. Compared with chips that can only pre-embed reaction reagents in the reaction chambers, this structural design of the present invention can add specific reaction reagents to the reaction chambers in the same detection unit according to the actual detection needs through the zigzag quantitative distribution channel, and achieve precise quantification.

[0028] (4) In a preferred embodiment of the centrifugal microfluidic high-throughput analytical chip provided by the present invention, when each detection unit is provided with a zigzag quantitative dispensing channel, the quantitative dispensing channels of each detection unit share a liquid storage chamber. Through this liquid storage chamber, specific reaction reagents can be added to the reaction chambers in each detection unit according to actual detection needs, and accurate quantification can be achieved through the quantitative chambers on the zigzag quantitative dispensing channels. In a further preferred design, the connection between the storage chamber and the sample inlet in each zigzag quantitative dispensing channel is set as an outward protrusion, which can assist in quantification.

[0029] (5) In a preferred embodiment of the centrifugal microfluidic high-throughput analysis chip provided by the present invention, the end of each detection unit furthest from the center of rotation is connected to the corresponding sealed gas chamber via a hydrophobic microchannel. During detection, after the reaction reagents enter the reaction chamber and undergo a mixing reaction, centrifugation continues. Some liquid will move along the hydrophobic microchannel between the reaction chamber and the sealed gas chamber towards the sealed gas chamber, causing the gas in the sealed gas chamber to be compressed. After reducing the rotation speed or stopping centrifugation, the compressed gas in the sealed gas chamber will release pressure and discharge the liquid that entered the hydrophobic microchannel. This alternating compression and release of the gas in the sealed gas chamber can accelerate the thorough mixing of the liquid in the reaction chamber and improve the reaction rate. In a further preferred design, the hydrophobic microchannel between the reaction chamber and the sealed gas chamber is zigzag-shaped, thereby increasing resistance.

[0030] (6) In the preferred embodiment of the centrifugal microfluidic high-throughput analysis chip provided by the present invention, the end of each detection unit away from the center of rotation is also connected to the corresponding temporary storage chamber through a capillary channel. During the detection process, after the reaction reagent enters the reaction chamber and undergoes a mixing reaction, centrifugation continues, and the liquid in the reaction chamber will enter the temporary storage chamber along the capillary channel. After the rotation speed is reduced or centrifugation is stopped, the liquid in the temporary storage chamber will return to the reaction chamber under the capillary force of the capillary channel. In this way, the liquid flows back and forth between the reaction chamber and the temporary storage chamber, which can accelerate the full mixing of the liquid in the reaction chamber and improve the reaction rate.

[0031] (7) In the preferred embodiment of the centrifugal microfluidic high-throughput analysis chip provided by the present invention, the middle region of each reaction chamber is hydrophilic while the other regions are output. During the detection process, after the reaction reagent enters the reaction chamber and undergoes a mixing reaction, centrifugation continues. The liquid in the reaction chamber will gather at the end away from the center of rotation under the action of centrifugal force. After the rotation speed is reduced or centrifugation is stopped, the liquid will return to the hydrophilic central region due to the action of surface tension. In this way, the liquid moves back and forth between the end away from the center of rotation and the central region in the reaction chamber, which can accelerate the full mixing of the liquid in the reaction chamber and improve the reaction rate. Attached Figure Description

[0032] Figure 1 is a schematic diagram of the centrifugal microfluidic high-throughput analysis chip structure provided in Embodiment 1 of the present invention;

[0033] Figure 2 is a schematic diagram of the detection unit structure in the centrifugal microfluidic high-throughput analysis chip provided in Embodiment 1 of the present invention;

[0034] Figure 3 is a schematic diagram of the centrifugal microfluidic high-throughput analysis chip structure provided in Embodiment 2 of the present invention;

[0035] Figure 4 is a schematic diagram of the detection unit structure in the centrifugal microfluidic high-throughput analysis chip provided in Embodiment 2 of the present invention;

[0036] Figure 5 is a schematic diagram of the centrifugal microfluidic high-throughput analysis chip structure provided in Embodiment 3 of the present invention;

[0037] Figure 6 is a schematic diagram of the detection unit structure in the centrifugal microfluidic high-throughput analysis chip provided in Embodiment 3 of the present invention;

[0038] Figure 7 is a schematic diagram of the centrifugal microfluidic high-throughput analysis chip structure provided in Embodiment 4 of the present invention;

[0039] Figure 8 is a schematic diagram of the detection unit structure in the centrifugal microfluidic high-throughput analysis chip provided in Embodiment 4 of the present invention;

[0040] Figure 9 is a schematic diagram of the centrifugal microfluidic high-throughput analysis chip structure provided in Embodiment 5 of the present invention;

[0041] Figure 10 is a schematic diagram of the detection unit structure in the centrifugal microfluidic high-throughput analysis chip provided in Embodiment 5 of the present invention.

[0042] Figure 11 is a schematic diagram of the centrifugal microfluidic high-throughput analysis chip structure provided in Embodiment 6 of the present invention;

[0043] Figure 12 is a schematic diagram of the detection unit structure in the centrifugal microfluidic high-throughput analysis chip provided in Embodiment 6 of the present invention;

[0044] Figure 13 is a schematic diagram of the centrifugal microfluidic high-throughput analysis chip structure provided in Embodiment 7 of the present invention;

[0045] Figure 14 is a schematic diagram of the detection unit structure in the centrifugal microfluidic high-throughput analysis chip provided in Embodiment 7 of the present invention;

[0046] Figure 15 is a schematic diagram of the centrifugal microfluidic high-throughput analysis chip structure provided in Embodiment 8 of the present invention;

[0047] Figure 16 is a schematic diagram of the detection unit structure in the centrifugal microfluidic high-throughput analysis chip provided in Embodiment 8 of the present invention;

[0048] Figure 17 shows the reaction chambers under different states in Embodiment 8 of the present invention;

[0049] In all the accompanying drawings, the same reference numerals are used to denote the same elements or structures, wherein:

[0050] 10- Rotation center, 11- Waste liquid chamber, 12- Air vent;

[0051] 20 - Reaction chamber; 21 - Central region of the reaction chamber; 22 - Region outside the central region of the reaction chamber;

[0052] 30 - Linear quantitative dispensing channel; 31 - First sample dispensing port; 32 - First quantitative chamber; 33 - Hydrophobic microchannel;

[0053] 40 - Quantitative sample dispensing unit, 41 - Independent sample dispensing port, 42 - Independent quantitative chamber, 43 - Independent waste liquid chamber, 44 - Capillary microchannel;

[0054] 50 - Zygmalion-shaped quantitative distribution channel; 51 - Second sample dispensing port; 52 - Second quantitative chamber; 53 - Hydrophobic microchannel;

[0055] 60 - Storage chamber; 61 - Shared sample dispensing port; 62 - Protrusion;

[0056] 70 - Sealed gas chamber; 72 - Hydrophobic microchannel;

[0057] 80 - Temporary liquid storage chamber, 81 - Capillary channel. Detailed Implementation

[0058] 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.

[0059] In this invention, the terms "first," "second," etc. (if present) in the invention and the accompanying drawings are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence.

[0060] This invention improves the distribution of chambers within the detection unit of a microfluidic chip and the connection method between chambers, so that during centrifugation, the microchannels connected to the quantitative chambers provide resistance matching the centrifugal force, thus achieving precise liquid quantification while facilitating chip fabrication. Specifically, in this embodiment, multiple reaction chambers in the same detection unit are distributed radially from the inside out, i.e., multiple reaction chambers are distributed on circumferences of different radii. At the same time, the quantitative chambers on the same quantitative distribution channel (linear or zigzag) in the detection unit are also distributed on circumferences of different radii, and the multiple reaction chambers and multiple quantitative chambers correspond sequentially along the direction from the rotation center to the edge. The connection, i.e., on the same quantitative dispensing channel, is such that the quantitative chamber farther from the rotation center is connected to a reaction chamber farther from the rotation center. Through this structural design, the hydrophobic microchannel between the quantitative chamber farther from the rotation center and the corresponding reaction chamber will also be longer, thereby providing greater resistance during centrifugation. Since the quantitative chamber farther from the rotation center will be subjected to greater centrifugal force during centrifugation, the structure designed in this invention does not require special design of the length and arrangement of the hydrophobic microchannels, so that the microchannel resistance experienced by the quantitative chamber can match the centrifugal force, effectively improving valve control stability, while also facilitating chip fabrication.

[0061] The chip provided by this invention is essentially a centrifugal microfluidic high-throughput analysis chip with radially extended detection units.

[0062] The chip provided by this invention can be equipped with multiple detection units, and each detection unit is equipped with multiple reaction chambers, thus enabling high-throughput analysis and detection.

[0063] In this invention, the hydrophobic microchannel possesses hydrophobic properties, either by being an inherently hydrophobic material or by hydrophobic modification. Examples of hydrophobic materials include polydimethylsiloxane (PDMS), toner, and wax films; the hydrophobic modification agent is a coating of one of the following on the microchannel surface: wax, silanizing agent, Teflon AF, CYTOP, and ECG1700.

[0064] In this invention, the number of linear quantitative dispensing channels can be set according to the types of reagents involved in the actual detection. It is easy to understand that, to avoid interference between linear quantitative dispensing channels, a maximum of two linear quantitative dispensing channels can be set in the same chip layer. When the number of linear quantitative dispensing channels is large, more chip layers are needed to accommodate them. Without loss of generality, the following embodiments will use two linear quantitative dispensing channels as an example for illustration.

[0065] The following is an example.

[0066] Example 1:

[0067] A centrifugal microfluidic high-throughput analysis chip, as shown in Figure 1, has a rotation center 10 and includes 6 detection units arranged around the rotation center.

[0068] As shown in Figure 2, the detection unit includes: a waste liquid chamber 11, two linear quantitative distribution channels 30, and four reaction chambers 20;

[0069] The four reaction chambers 20 are distributed radially from the inside to the outside.

[0070] The linear quantitative distribution channel 30 extends radially, and its inner and outer ends form the beginning and end, respectively. The beginning end is provided with a first sample inlet 31, and the end is connected to the waste liquid chamber 11. The linear quantitative distribution channel 30 is provided with four first quantitative chambers 32. The four first quantitative chambers 32 are respectively connected to the four reaction chambers 20 in sequence from the rotation center to the edge through hydrophobic microchannels 33.

[0071] In the interconnected first quantitative chamber 32 and reaction chamber 20, the distance of reaction chamber 20 from the rotation center is greater than the distance of the first quantitative chamber 32 from the rotation center.

[0072] As shown in Figures 1 and 2, in this embodiment, the waste liquid chamber 11 is also connected to the air vent 12 to balance the air pressure inside the waste liquid chamber.

[0073] The centrifugal microfluidic high-throughput analytical chip provided in this embodiment can pre-embed a reaction chamber containing a reagent (such as powder, which allows for the mixing of the quantitatively dispensed liquid reagent and powder). Two other reagents can be added through two linear quantitative dispensing channels. Precise quantitative dispensing of the liquid is achieved through low-speed centrifugation or simple manual injection. Finally, high-speed centrifugation completes the mixing of the different liquids. Specifically, when using low-speed centrifugation (e.g., 300 rpm) or simple manual injection, the liquid enters the quantitative chamber but does not breach the hydrophobic microchannels. Excess liquid flows into the waste liquid pool through the linear quantitative dispensing channels. After quantification, high-speed centrifugation (e.g., 1000 rpm) is used. The quantified liquid enters the reaction chamber sequentially from the center of rotation towards the edge for mixing. Since some liquid remains at the reagent dispensing port during low-speed centrifugation, this excess liquid enters the quantitative chamber closest to the beginning during high-speed centrifugation without affecting the quantitative dispensing in the reaction chamber. Therefore, as a preferred embodiment, in this embodiment, the quantitative chamber closest to the beginning of each linear quantitative dispensing channel is used as a sacrificial chamber to accommodate excess reagents or excess samples generated during centrifugation.

[0074] In addition, there are various mixing methods. For example, preliminary mixing can be achieved by controlling the order in which reagents or samples enter; mixing can also be accomplished by controlling the acceleration of the centrifugation speed and the continuous change of the centrifugation direction. For example, after the liquid enters the reaction chamber, the mixing of the reaction reagents can be accelerated and the reaction efficiency improved by reversing the direction at a low speed (e.g., 500 rpm).

[0075] Referring to Figures 1 and 2, in this embodiment, within the same detection unit, the first quantitative chambers on each linear quantitative distribution channel, excluding the sacrificial chamber, have the same volume. Therefore, the centrifugal microfluidic high-throughput analysis chip provided in this embodiment can be used for high-throughput screening of antibiotics with equivalent quantitative properties. Specifically, bacterial suspensions containing different types of antibiotics are pre-embedded in the reaction chamber, with at least two types of each antibiotic selected. Two concentrations of the same antibiotic are pre-embedded in adjacent reaction chambers at different concentrations. The pre-embedded antibiotics can be in the form of dry powder, coated paper, or hydrogel co-lyophilized, depending on actual needs. Buffer and acid-base indicator are added to the first sample inlet of the two linear quantitative dispensing channels in each detection unit, respectively. After centrifugation at 300 rpm, the bacterial suspension, buffer, and acid-base indicator enter the quantitative chamber along their respective reagent quantitative dispensing channels. Due to the hydrophobic microchannels, liquid does not enter the reaction chamber; excess waste liquid enters the waste liquid chamber. The rotation speed is increased to 1000 rpm, and the liquid in the quantitative chamber breaks through the hydrophobic microchannels and enters the reaction chamber to mix with the pre-embedded antibiotics. Low-speed forward and reverse rotation at 500 rpm accelerates the mixing of reaction reagents. After co-incubation at 37 degrees Celsius for a certain period of time, the results can be read, thereby achieving high-throughput screening of antibiotics.

[0076] Referring to Figures 1 and 2, in this embodiment, the chip has a disk-like structure. Optionally, in this embodiment, the chip is designed as a double-layer structure, consisting of a microchannel layer and a reagent layer. The hydrophobic microchannel is located in the channel layer, while the remaining chamber structure and the first sample dispensing port are located in the reagent layer.

[0077] The centrifugal microfluidic high-throughput analysis chip provided in this embodiment can be processed from materials such as PMMA, PC, PP, and PDMS, and the processing methods include, but are not limited to, CNC machining, laser engraving, soft lithography, 3D printing, and injection molding. For example, since the main structure of the disk-type chip consists of a microfluidic channel layer and a reagent layer, when the chip is processed using CNC technology, only the two-layer structure needs to be bonded; when processed using laser engraving technology, only the top and bottom cover plates need to be added.

[0078] It should be noted that in this embodiment, the number of detection units in the chip, the number of linear quantitative distribution channels, the number of reaction chambers, the number of chip layers, etc., are merely illustrative descriptions and should not be construed as the sole limitation of the present invention. In other detection scenarios, these parameters can be flexibly set according to actual detection needs.

[0079] Example 2:

[0080] A centrifugal microfluidic high-throughput analysis chip, the structure of which is shown in Figures 3 and 4. This embodiment is similar to Embodiment 1 above, except that, as shown in Figures 3 and 4, in the two linear quantitative dispensing channels 30 of the same detection unit, the volume of the first quantitative chamber on one channel gradually decreases from the beginning to the end, excluding the sacrificial chamber; while the volume of the first quantitative chamber on the other channel gradually increases from the beginning to the end, excluding the sacrificial chamber. Through this structural design, this embodiment can achieve liquid quantification based on concentration gradients.

[0081] In this embodiment, the specific implementation of the remaining structures can be referred to the description in Embodiment 1 above, and will not be repeated here.

[0082] It is easy to understand that when the detection unit is equipped with more linear quantitative dispensing channels, more concentration gradients can be constructed for the added detection reagents by setting the volume of the first quantitative chamber, which greatly increases the detection diversity.

[0083] Example 3:

[0084] A centrifugal microfluidic high-throughput analysis chip, the structure of which is shown in Figures 5 and 6, is similar to the above-described embodiment 2. The difference is that, as shown in Figures 5 and 6, in this embodiment, each reaction chamber in the detection unit is further provided with a quantitative sample addition unit 40 on the side near the rotation center. The quantitative sample addition unit 40 includes: an independent sample addition port 41, and an independent quantitative chamber 42 and an independent waste liquid chamber 43 connected to the independent sample addition port 41. Each independent quantitative chamber 42 is connected to the corresponding reaction chamber 20 through a capillary microchannel 44.

[0085] As shown in Figures 5 and 6, in this embodiment, the independent waste liquid chamber 43 in the independent sample addition unit 40 is also connected to an air vent to balance the air pressure in the independent waste liquid chamber.

[0086] When adding reagents to the reaction chamber using a pre-embedded method, the type of reagent cannot be changed, and it is difficult to achieve precise quantification of the reagent. This embodiment, by setting up corresponding independent sample addition units for each reaction chamber, allows for the addition of the same reagent to multiple reaction chambers or the addition of different reagents to different reaction chambers, depending on the specific testing requirements.

[0087] In this embodiment, the independent sample dispensing port and the reaction chamber are connected to the independent waste liquid chamber by a curved capillary microchannel, which increases the resistance of the liquid during centrifugation, slows down the flow of liquid reaction reagents, and improves reagent utilization.

[0088] In specific application scenarios, in this embodiment, each detection unit can be set with two variables: First, for each detection unit, if the same reagent is added through an independent sample dispensing port, two different reagents can be added to the two outer linear dispensing channels to construct a liquid concentration gradient; Second, for each detection unit, if different reagents are added through the independent sample dispensing ports on the inner side, the same reagent or different reagents can be added to the two linear dispensing channels, which greatly improves the diversity of detection reagents.

[0089] In this embodiment, the specific implementation of the remaining structures and the control method when the chip is working can be referred to the descriptions in Embodiments 1 and 2 above, and will not be repeated here.

[0090] Example 4:

[0091] A centrifugal microfluidic high-throughput analysis chip, the structure of which is shown in Figures 7 and 8. This embodiment is similar to Embodiment 2 above, except that in this embodiment, each detection unit further includes: a zigzag quantitative dispensing channel 50 extending from the rotation center to the edge;

[0092] The inner and outer ends of the zigzag quantitative distribution channel 50 constitute its head and tail ends, respectively. The head end is provided with a second sample inlet 51, and the tail end is connected to the waste liquid chamber 11. The zigzag quantitative distribution channel 50 is provided with four second quantitative chambers 52. The four second quantitative chambers 52 are respectively connected to the four reaction chambers 20 in sequence from the rotation center to the edge through hydrophobic microchannels 53.

[0093] In the interconnected second quantitative chamber 52 and reaction chamber 20, the distance of reaction chamber 20 from the rotation center is greater than the distance of the second quantitative chamber 52 from the rotation center.

[0094] The zigzag quantitative distribution channel allows for the addition of specific reaction reagents to the reaction chambers within the same detection unit according to actual detection needs, achieving precise quantification.

[0095] In this embodiment, the specific implementation of the remaining structures can be referred to the descriptions in Embodiments 1 and 2 above, and will not be repeated here.

[0096] Example 5:

[0097] A centrifugal microfluidic high-throughput analysis chip, the structure of which is shown in Figures 9 and 10. This embodiment is similar to Embodiment 4 above, except that in this embodiment, the chip further includes a storage chamber 60 disposed between the rotation center and the four detection units, which is provided with a common sample dispensing port 61, and the second sample dispensing port 51 of the zigzag quantitative dispensing channel 50 in the detection unit is connected to the storage chamber 60.

[0098] After adding the reaction reagent to the storage chamber through a shared sample dispensing port and centrifuging, the same reaction reagent can be added to the reaction chamber within the same detection unit through the zigzag quantitative dispensing channel in each detection unit.

[0099] Referring to Figures 9 and 10, in this embodiment, the connection between the storage chamber 60 and the second sample application port 51 is configured as an outward protrusion 62, which can be used to assist in quantitative analysis.

[0100] For the specific implementation of the remaining structures in this embodiment, please refer to the description in Embodiment 4 above.

[0101] The specific implementation plan for generating antibiotic concentration gradients using this chip for microbial antimicrobial susceptibility testing is described below:

[0102] Bacterial suspension is added to a common sampling port, and a single sampling can be used for high-throughput screening of multiple antibiotics, simplifying the operation steps; antibiotic solution and buffer solution with acid-base indicator are added to the first sampling port of the two linear quantitative dispensing channels in each detection unit, respectively. Different types of antibiotic solutions are added to different detection units, which can realize high-throughput screening of multiple antibiotics.

[0103] The chip center is fixed on the centrifuge shaft and centrifuged at low speed (300 rpm). The three solutions enter the quantitative chamber through the quantitative distribution channel respectively. Due to the hydrophobic microchannel, the resistance of the liquid in the quantitative chamber is greater than the centrifugal force under these conditions, and the solution temporarily stays in the quantitative chamber.

[0104] Within the same detection unit, in a linear quantitative dispensing microchannel, excluding the sacrificial chamber, the volume of the quantitative chamber gradually decreases from the beginning to the end; in another linear reagent quantitative dispensing channel, excluding the sacrificial chamber, the volume of the quantitative chamber gradually increases from the beginning to the end; in a zigzag reagent quantitative dispensing channel, the volume of the quantitative chamber remains constant from the beginning to the end. This design can achieve equal quantitative dispensing of bacterial suspensions and gradient quantitative dispensing of antibiotic solutions.

[0105] Increasing the rotation speed to 1200 rpm reduces the resistance experienced by the liquid in the metering chamber to less than the effect of centrifugal force. Simultaneously, guided by the hydrophobic microchannel, the liquid in the three metering chambers corresponding to each reaction chamber enters the reaction chamber, achieving mixing of the reaction reagents.

[0106] The reaction is accelerated by rotating the forward and reverse directions at 500 rpm to mix the reagents. After co-culturing at 37 degrees Celsius for a certain period of time, the results can be read, thus enabling the detection of microbial drug susceptibility.

[0107] Example 6:

[0108] A centrifugal microfluidic high-throughput analysis chip, the structure of which is shown in Figures 11 and 12. This embodiment is similar to Embodiment 1 above, except that in this embodiment, the chip further includes: four sealed gas chambers 70 corresponding one-to-one with the four reaction chambers 20, and the end of each reaction chamber 20 away from the center of rotation is connected to the corresponding sealed gas chamber 70 through a hydrophobic microchannel 72.

[0109] Based on this structural design, when the centrifugal microfluidic high-throughput analysis chip provided in this embodiment is used for detection, after the reaction reagents enter the reaction chamber and undergo a mixing reaction, centrifugation continues. Some of the liquid will move along the hydrophobic microchannel between the reaction chamber and the sealed gas chamber towards the sealed gas chamber, causing the gas in the sealed gas chamber to be compressed. After reducing the speed or stopping centrifugation, the compressed gas in the sealed gas chamber will release pressure and discharge the liquid that entered the hydrophobic microchannel. In this way, the gas in the sealed gas chamber is alternately compressed and released, which can accelerate the thorough mixing of the liquid in the reaction chamber and improve the reaction rate.

[0110] As shown in Figures 11 and 12, in this embodiment, the hydrophobic microchannel 72 connecting the sealed gas chamber 70 and the reaction chamber 20 is of a zigzag shape, which increases resistance. In this embodiment, the specific implementation of the remaining structures can be referred to the description in Embodiment 1 above, and will not be repeated here.

[0111] Example 7:

[0112] A centrifugal microfluidic high-throughput analysis chip, the structure of which is shown in Figures 13 and 14. This embodiment is similar to Embodiment 1 above, except that in this embodiment, the chip further includes four temporary liquid storage chambers 80 corresponding to the four reaction chambers 20, and the end of each reaction chamber 20 away from the center of rotation is connected to the corresponding temporary liquid storage chamber 80 through a capillary channel 81.

[0113] The centrifugal microfluidic high-throughput analysis chip provided in this embodiment utilizes the wetting properties of liquids to accelerate the mixing of liquids within the reaction chamber. When the chip is used for detection, after the reagents enter the reaction chamber and undergo a mixing reaction, centrifugation continues. The liquid in the reaction chamber flows along capillary channels into a temporary reservoir. When the rotation speed is reduced or centrifugation is stopped, the liquid in the temporary reservoir returns to the reaction chamber under the capillary force of the capillary channels. This reciprocating flow of liquid between the reaction chamber and the temporary reservoir accelerates the thorough mixing of the liquid in the reaction chamber and increases the reaction rate.

[0114] Example 8:

[0115] A centrifugal microfluidic high-throughput analysis chip, the structure of which is shown in Figures 15 and 16. This embodiment is similar to Embodiment 1 above, except that in this embodiment, the central region 21 of the reaction chamber 20 is hydrophilic, while the region 22 outside the central region is hydrophobic. Specifically, in this embodiment, the central region of the reaction chamber is made of a hydrophilic material, while the region outside the central region is made of a hydrophobic material, thereby giving different regions different hydrophilicities.

[0116] When the chip provided in this embodiment is used for detection, after the reaction reagents enter the reaction chamber and undergo a mixing reaction, centrifugation continues. Under the action of centrifugal force, the liquid in the reaction chamber will gather at the end away from the center of rotation, as shown in the left half of Figure 17. After reducing the speed or stopping centrifugation, due to the effect of surface tension, the liquid will return to the hydrophilic central region, as shown in the right half of Figure 17. In this way, the liquid moves back and forth between the end away from the center of rotation and the central region in the reaction chamber, which can accelerate the thorough mixing of the liquid in the reaction chamber and improve the reaction rate.

[0117] In this embodiment, the specific implementation of the remaining structures can be referred to the description in Embodiment 1 above, and will not be repeated here.

[0118] 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 centrifugal microfluidic high-throughput analysis chip, wherein a rotation center is provided, characterized in that, include: The system comprises M detection units arranged around the rotation center; each detection unit includes: a waste liquid chamber, N linear quantitative dispensing channels, and K reaction chambers; the K reaction chambers are arranged radially from the inside to the outside; each linear quantitative dispensing channel extends radially, with its inner and outer ends forming a head and a tail, respectively, the head end having a first sample inlet and the tail end communicating with the waste liquid chamber; K first quantitative chambers are arranged on the linear quantitative dispensing channels; each of the K first quantitative chambers is connected to the K reaction chambers sequentially from the rotation center to the edge via hydrophobic microchannels; in the interconnected first quantitative chambers and reaction chambers, the distance of the reaction chamber from the rotation center is greater than the distance of the first quantitative chamber from the rotation center; the detection unit further includes: a zigzag quantitative dispensing channel extending from the rotation center to the edge; the zigzag... The inner and outer ends of the linear quantitative distribution channel constitute its head and tail, respectively. The head is provided with a second sample inlet, and the tail is connected to the waste liquid chamber. K second quantitative chambers are provided on the linear quantitative distribution channel. The K second quantitative chambers are sequentially connected to the K reaction chambers along the direction from the rotation center to the edge via hydrophobic microchannels. In the interconnected second quantitative chambers and reaction chambers, the distance from the reaction chamber to the rotation center is greater than the distance from the second quantitative chamber to the rotation center. The detection unit also includes K sealed gas chambers corresponding one-to-one with the K reaction chambers, and the end of each reaction chamber away from the rotation center is connected to the corresponding sealed gas chamber via a hydrophobic microchannel. The central region of the reaction chamber is hydrophilic, and the region outside the central region is hydrophobic. Wherein, M, N, and K are all positive integers, and K > 1.

2. The centrifugal microfluidic high-throughput analysis chip as described in claim 1, characterized in that, In the detection unit, each reaction chamber is further provided with a quantitative sample addition unit on the side near the rotation center; the quantitative sample addition unit includes: an independent sample addition port, and an independent quantitative chamber and an independent waste liquid chamber connected to the independent sample addition port; each independent quantitative chamber is connected to the corresponding reaction chamber through a capillary microchannel.

3. The centrifugal microfluidic high-throughput analysis chip as described in claim 1, characterized in that, Also includes: The storage chamber is located between the rotation center and the M detection units, and has a common sample dispensing port. The second sample dispensing port of the zigzag quantitative dispensing channel in the detection unit is connected to the storage chamber.

4. The centrifugal microfluidic high-throughput analysis chip as described in claim 3, characterized in that, The connection between the storage chamber and the second sample application port is designed as an outward protrusion.

5. The centrifugal microfluidic high-throughput analysis chip according to any one of claims 1 to 4, characterized in that, The hydrophobic microchannel connecting the sealed gas chamber and the reaction chamber is zigzag-shaped.

6. The centrifugal microfluidic high-throughput analysis chip according to any one of claims 1 to 4, characterized in that, The detection unit further includes K temporary liquid storage chambers that correspond one-to-one with the K reaction chambers, and the end of each reaction chamber away from the rotation center is connected to the corresponding temporary liquid storage chamber through a capillary channel.

7. The centrifugal microfluidic high-throughput analysis chip according to any one of claims 1 to 4, characterized in that, In each linear quantitative dispensing channel, the quantitative chamber closest to the head is the sacrificial chamber.

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