Microfluidic chip and microfluidic chip operating system

Abstract

To provide a microfluidic chip and microfluidic chip operating system.SOLUTION: A microfluidic chip includes: a sample injection groove, a processing chamber, a first filter element, a chromatography column, a liquid flow path system, and a detection region that are sequentially connected; a first valve element interposed between the sample injection groove and the processing chamber; a second valve element interposed between the processing chamber and the first filter element; and an actuator element provided above the processing chamber, including a drive membrane, and configured to generate a vortex in the processing chamber and to control the pressure inside the processing chamber.SELECTED DRAWING: Figure 7

Description

[Technical Field]

[0001] The present disclosure relates to a microfluidic chip for extracting compounds from a sample and an associated microfluidic chip operating system. [Background technology]

[0002] When detecting substances contained in biological or chemical samples, the samples are often complex mixtures, so it is often necessary to first isolate and purify specific compounds. Conventional extraction and isolation processes involve first extracting the sample with an extraction solvent, followed by isolation and purification to obtain the compounds for subsequent detection, such as determining the types and properties of the compounds in the sample. However, conventional extraction and isolation procedures often require the collection of large amounts of sample material, as well as the use of large amounts of reagents, consumables, and labor. Summary of the Invention [Problem to be solved by the invention]

[0003] In view of the above problems, an object of the present disclosure is to provide a microfluidic chip and a microfluidic chip operating system for integrating and completing the operations of extracting and separating compounds in a sample into a microfluidic chip. [Means for solving the problem]

[0004] Some embodiments of the present disclosure provide a microfluidic chip including a sample injection channel, a processing chamber, a first filter element, a chromatography column, a liquid flow path system, and a detection region, which are connected in sequence; a first valve element provided between the sample injection channel and the processing chamber; a second valve element provided between the processing chamber and the first filter element; and an actuator element provided above the processing chamber, including a drive membrane, and configured to generate a vortex flow in the processing chamber and control the pressure in the processing chamber.

[0005] In some embodiments, the first valve element includes a first valve column having an opening dimension in the range of about 5 micrometers to about 30 micrometers.

[0006] In some embodiments, the lower surface of the drive membrane is an uneven structure.

[0007] In some embodiments, the microfluidic chip further comprises a heating element disposed below the processing chamber.

[0008] In some embodiments, the microfluidic chip further includes a weighing element disposed below the processing chamber.

[0009] In some embodiments, the microfluidic chip further comprises a second filter element interposed between the chromatography column and the liquid flow path system.

[0010] In some embodiments, the detection region comprises a plurality of first micropores.

[0011] In some embodiments, the detection region comprises multiple sub-detection regions, each containing at least one micropore.

[0012] In some embodiments, the detection region includes a first sub-detection region including a plurality of first micropores, a second sub-detection region including a plurality of second micropores, a third sub-detection region including a plurality of third micropores, and a fourth sub-detection region including a plurality of fourth micropores.

[0013] In some embodiments, the liquid flow path system includes a main flow path and a plurality of branch flow paths. In some embodiments, the main flow path is connected to a plurality of primary branch flow paths. In some embodiments, the plurality of primary branch flow paths are each connected to a plurality of secondary branch flow paths.

[0014] In some embodiments, the first plurality of micropores are provided with cells, antibodies, signal sensing elements, or combinations thereof.

[0015] In some embodiments, the microfluidic chip further includes a reagent addition / sampling channel connected to the chromatography column, and a third valve element configured to control the liquid flow direction and opening / closing of the reagent addition / sampling channel.

[0016] In some embodiments, the microfluidic chip further comprises a waste flow path in communication with the chromatography column, and a fourth valve element configured to control opening and closing of the waste flow path.

[0017] In some embodiments, in the microfluidic chip, the first valve element, the second valve element, and the actuator element are gas pressure controlled.

[0018] In some embodiments, the microfluidic chip is a stack structure including a substrate layer, a first flow path layer above the substrate layer and having a plurality of recessed grooves defined therein for liquid flow, and a second flow path layer above the first flow path layer and having a plurality of recessed grooves defined therein for gas flow.

[0019] In some embodiments, the first flow path layer includes a plurality of first openings, and the second flow path layer includes a plurality of second openings each corresponding to the plurality of first openings, and the plurality of first openings and the plurality of second openings define a plurality of liquid storage regions.

[0020] In some embodiments, the first filter of the first filter element is provided in a first filtration chamber consisting of a first filter groove in the first flow path layer and a second filter groove in the second flow path layer, and the second filter of the second filter element is provided in a second filtration chamber consisting of a third filter groove in the first flow path layer and a fourth filter groove in the second flow path layer.

[0021] In some embodiments, the microfluidic chip further comprises a heated weighing component disposed within the substrate layer such that it is at the bottom of the process chamber.

[0022] In some embodiments, in a microfluidic chip, the actuator element includes a drive membrane and a drive control groove above the drive membrane.

[0023] In some embodiments, the first flow path layer includes a process chamber groove and a drive membrane overlying the process chamber groove, the position of the process chamber groove corresponding to the process chamber, and the second flow path layer includes a drive control groove.

[0024] In some embodiments, the actuator element further includes a drive control groove above the drive membrane, the processing chamber and the drive membrane being provided in the first flow path layer, and the drive control groove being provided in the second flow path layer.

[0025] In some embodiments, in the microfluidic chip, the second flow channel layer further includes a driving gas channel communicating with the driving control groove, and a driving gas vent hole communicating with the driving gas channel.

[0026] In some embodiments, in the microfluidic chip, the first valve element includes: a first valve column located between the sample injection groove and the processing chamber and provided in the first flow path layer; a first elastic membrane located higher than the first valve column and provided in the first flow path layer; and a first valve control groove located above the first valve column and the first elastic membrane and provided in the second flow path layer.

[0027] In some embodiments, in the microfluidic chip, the second flow path layer further includes a first valve gas flow path communicating with the first valve control groove, and a first valve gas vent communicating with the first valve gas flow path.

[0028] In some embodiments, in the microfluidic chip, the chromatography column is disposed in the second flow path layer.

[0029] In some embodiments, in a microfluidic chip, a first flow path layer includes a first sample injection opening, and a second flow path layer includes a second sample injection opening, and the positions of the first sample injection opening and the second sample injection opening correspond to the sample injection groove.

[0030] In some embodiments, the detection region includes a plurality of first micro-holes, the first flow path layer includes a plurality of first micro-through-holes, and the second flow path layer includes a plurality of second micro-through-holes, and these first micro-holes correspond to these first micro-through-holes and these second micro-through-holes, respectively.

[0031] In some embodiments, in the microfluidic chip, the sample injection channel has a volume larger than the volume of the processing chamber.

[0032] Some embodiments of the present disclosure provide a microfluidic chip operating system including a microfluidic chip as described in the above and following embodiments, an actuator element driving module configured to control the raising and lowering of an actuator element of the microfluidic chip, and a valve driving module configured to control the opening and closing of a first valve element and a second valve element.

[0033] In some embodiments, in the microfluidic chip operating system, the actuator element driving module is configured to pneumatically control the deformation, for example, lifting and lowering, of the driving membrane of the actuator element.

[0034] In some embodiments, in the microfluidic chip operating system, the valve driving module is configured to pneumatically control the opening and closing of the first valve element and the second valve element.

[0035] In some embodiments, the microfluidic chip operating system further includes a waste collection module configured to collect effluent from the chromatography column and a reagent addition module configured to add reagents to the inflow / outflow channels.

[0036] In some embodiments, the microfluidic chip operating system further comprises a detection instrument for detecting fluid from said chromatography column or detection region.

[0037] In some embodiments, the detection instrument is a mass spectrometer or a chromatography device, such as a coupled plasma mass spectrometer, a liquid phase chromatography device, or a gas phase chromatography device. [Brief explanation of the drawings]

[0038] The various aspects of the present disclosure are best understood when read in conjunction with the drawings, which illustrate, by way of example, the principles of the present disclosure. It should be noted that, in accordance with standard industry practices, various features may not be drawn to scale. In fact, the sizes of various features may be arbitrarily increased or decreased for clarity of discussion. To more clearly illustrate the above and other objects, features, advantages, and embodiments of the present disclosure, the drawings are set forth as follows: [Figure 1] FIG. 1 is a perspective view of a microfluidic chip according to some embodiments. [Figure 2] 1A-1C are top views of a microfluidic chip according to some embodiments. [Figure 3] FIG. 1 is an exploded view of a microfluidic chip according to some embodiments. [Figure 4] FIG. 1 is a top view of a substrate layer of a microfluidic chip according to some embodiments. [Figure 5] FIG. 2 is a top view of a first flow channel layer of a microfluidic chip according to some embodiments. [Figure 6] FIG. 1B is a top view of a second flow channel layer of a microfluidic chip according to some embodiments. [Figure 7] FIG. 1 is a layout diagram of a stack structure of a microfluidic chip according to some embodiments. [Figures 8A-8C] 8A-8C are cross-sectional views of the microfluidic chip of FIG. 7 along lines AB and BC during extraction, according to some embodiments. [Figure 9A] FIG. 10 is a bottom view of a driving membrane according to some embodiments. [Figure 9B] 9B is a cross-sectional view of the driving membrane of FIG. 9A along line FF' according to some embodiments. [Figure 9C]FIG. 10 is a bottom view of a driving membrane according to another embodiment. [Figure 10] FIG. 8 is a cross-sectional view of the microfluidic chip of FIG. 7 taken along line DD'. [Figure 11] FIG. 8 is a cross-sectional view of the microfluidic chip of FIG. 7 taken along line EE'. [Figure 12] FIG. 1 is a schematic diagram of a microfluidic chip operating system according to some embodiments. [Figure 13] 1 is a flowchart of a method of using a microfluidic chip according to some embodiments. DETAILED DESCRIPTION OF THE INVENTION

[0039] The following disclosure provides many different embodiments or examples for implementing different features of the provided subject matter. Below, specific embodiments of components and arrangements are presented to simplify the disclosure. Of course, these are merely examples and are not limiting. For example, in the following description, "forming a first feature above or on a second feature" may include an embodiment in which the first feature and the second feature are formed in direct contact with each other, or an embodiment in which an additional feature may be formed between the first feature and the second feature, and thus the first feature and the second feature may not be in direct contact with each other. Furthermore, the disclosure may repeat reference numerals and / or letters in various examples. Such repetition is for purposes of brevity and clarity, and the repetition itself does not imply a relationship between the respective embodiments and / or configurations discussed.

[0040] It should be noted that spatially relative terms such as "below," "lower than," "below," "higher than," "above," "on," "top," "above," and similar terms may be used herein to indicate the relationship of one element or feature to another element or feature shown in the figures. In addition to the orientation shown, the spatially relative terms are intended to cover different orientations of the device during use or operation. The device may be oriented in other ways (rotated 90 degrees or in another orientation) and the spatially relative descriptors used herein may be interpreted accordingly.

[0041] As used in this disclosure, terms such as "first," "second," etc., are used to modify elements and do not themselves imply any preceding number for this (or these) elements, nor do they imply any ordering of one element from another, or any ordering in a manufacturing process. The use of these numbers is only used to clearly distinguish one named element from another named element. The claims and the specification may not use the same number terms, and accordingly, a first element in the specification may be a second element in the claims.

[0042] Microfluidics refers to the science and technology of systems that process or manipulate microfluids using microchannels. Due to their miniaturization and integration, microfluidic devices are commonly called microfluidic chips.

[0043] Biological or chemical materials contain complex mixtures. When attempting to isolate active ingredients with specific effects, for example, by extracting secondary metabolites (e.g., compounds of the types flavonoids, polysaccharides, volatile oils, quinones, terpenes, saponins, alkaloids, pigments, coumarins, cardiac glycosides, and phenolic acids) from plants and screening for specific compounds with therapeutic effects against diseases, processes such as separation, purification, refinement, concentration, and drying must be performed, followed by cell or animal experiments. However, these processes often require large amounts of raw materials, reagents, consumables, labor, and time. In contrast, in some embodiments of the present disclosure, multiple operations, such as sample extraction and compound separation, are integrated into a single microfluidic chip, and the separated compounds can be directly used for detection. In some embodiments, detection, such as detection of cellular reactions, may be performed on the microfluidic chip, allowing the operations of extraction, compound separation, and screening for active ingredients to be completed on a single chip.

[0044] 1 to 7, which illustrate a microfluidic chip according to some embodiments. FIG. 1 is a perspective view of the microfluidic chip 10, FIG. 2 is a top view of the microfluidic chip 10, FIG. 3 is an exploded view of the microfluidic chip 10, FIGS. 4 to 6 are top views of each layer of the microfluidic chip 10, and FIG. 7 is a top layout diagram of each layer of the microfluidic chip 10. The microfluidic chip 10 includes a sample injection channel 410, a first valve element 420, a processing chamber 430, a second valve element 440, a first filter element 510, a chromatography column 520, a second filter element 530, a liquid flow path system 540, and a detection region 550, which are sequentially connected to each other. The arrangement of each of the above-mentioned connected components will be used below. After the sample and extraction solvent are introduced into the sample injection channel 410, the extraction solution can flow into the processing chamber 430 by opening the first valve element 420, and can also flow into the first filter element 510 by opening the second valve element 440. The extraction solution then enters the chromatography column 520, and compounds that have an affinity with the filler in the chromatography column 520 remain in the chromatography column 520. After being eluted with the eluent, the separated compounds can flow out, pass through the second filter element 530, and then enter the liquid flow system 540. The separated compounds can then be transported to the multiple micropores in the detection region for subsequent detection of several properties.

[0045] The microfluidic chip 10 further includes an actuator element 470 provided above the processing chamber 430. The actuator element 470 is configured to drive the flow of fluid within the microfluidic chip 10. The actuator element 470 includes a driving membrane 280, which is an elastic thin film, and when deformation such as rising and falling occurs, pressure is generated within the processing chamber 430, allowing the fluid to flow into or out of the processing chamber 430. In other words, the pressure within the processing chamber 430 can be controlled by the deformation of the driving membrane 280.

[0046] After the sample and extraction solvent are placed in the sample injection channel 410, the first valve element 420 is opened, the second valve element 440 is closed, and the actuator element 470 controls the driving membrane 280 to repeatedly raise and lower, thereby repeatedly drawing in and expelling the liquid in the sample injection channel 410 from the processing chamber 430. This generates a vortex, accelerating the mixing of the sample with the extraction solution and the release and dissolution of components in the sample into the extraction solvent. In other embodiments, the sample may be pre-treated, for example, with an ultrasonic device, to disrupt the cells in the sample more quickly. Because the pressure generated by the actuation of the driving membrane 280 requires multiple drawing and expelling of liquid into and from the processing chamber 430, the volume of the sample injection channel 410 may be larger than the volume of the processing chamber 430. In some embodiments, the volume of the sample injection channel 410 is at least five times or at least ten times the volume of the processing chamber 430.

[0047] Furthermore, because the liquid in the sample injection channel 410 must pass through the first valve element 420 before entering the processing chamber 430, any particles in the extraction solution larger than the opening dimension OD1 (see FIG. 8B ) of the first valve column 290 of the first valve element 420 cannot enter the processing chamber 430. Thus, the first valve element 420 provides an additional filtering effect, preventing tissue residue and debris in the sample from entering the processing chamber 430. In some embodiments, the opening dimension OD1 of the first valve column 290 of the first valve element 420 can be in the range of about 5 micrometers to about 30 micrometers.

[0048] In some embodiments, after the components to be extracted are substantially dissolved in the extraction solvent, first valve element 420 is closed, second valve element 440 is opened, and the extraction solution is forced to flow into first filter element 510 under pressure from actuation of drive membrane 280. First filter element 510 includes a first filtration chamber 512 and a first filter 514 located within first filtration chamber 512 for filtering small residues or impurities in the extraction solution. Actuation of drive membrane 280 then forces the extraction solution into chromatography column 520 under pressure. After chromatographic extraction is complete, the effluent passes through second filter element 530. Second filter element 530 includes a second filtration chamber 532 and a second filter 534 located within second filtration chamber 532 for filtering beads from chromatography column 520 that may appear in the effluent.

[0049] 1 to 7, the microfluidic chip 10 further includes a reagent addition / sampling channel 260 and a third valve element 450. The third valve element 450 is configured to control the liquid flow direction and opening / closing of the reagent addition / sampling channel 260. Optionally, after the eluent containing the compounds flows out of the chromatography column 520, the flowing liquid may be introduced into a detection device, such as a UV absorbance meter, a liquid-phase chromatography device, a gas-phase chromatography device, or a coupled plasma mass spectrometer, via the reagent addition / sampling channel 260 to measure the UV absorbance values ​​of the separated compounds or signals related to the components.

[0050] After leaving the chromatography column 520, the eluent containing the compound enters a plurality of micropores 554 in the detection region 550 via the liquid flow path system 540. Optionally, reagents required for detection may be added to the detection region 550 via the reagent addition / sampling channel 260. In some embodiments, to observe cellular responses to the same target compound at different concentrations, a dilution solution may be added to the micropores in the detection region via the reagent addition / sampling channel 260. The dilution solution may be, for example, phosphate buffered saline (PBS) or a cell culture medium.

[0051] The microfluidic chip 10 further includes a waste flow path 270 and a fourth valve element 460. The fourth valve element 460 is configured to control the opening and closing of the waste flow path 270. In some embodiments, the initial eluent or eluents of other stages passing through the chromatography column 520 do not contain the target compound, so the initial eluent or eluents of other stages are directed into the microfluidic chip 10 via the waste flow path 270 and then flow into an external waste channel (724, see FIG. 12 ).

[0052] 7, the liquid flow path system 540, the reagent addition / sampling flow path 260, and the waste flow path 270 are all connected to the second filtration chamber 532 of the second filter element 530. When the liquid outflow chromatography column 520 reaches the second filtration chamber 532 of the second filter element 530, the third valve element 450 and the fourth valve element 460 are controlled to direct the liquid to flow into one of the liquid flow path system 540, the reagent addition / sampling flow path 260, and the waste flow path 270.

[0053] 1-7 , the liquid flow path system 540 includes a main flow path 542, a plurality of primary branch flow paths 544, and a plurality of secondary branch flow paths 546. The main flow path 542 branches into a plurality of primary branch flow paths 544. Each of the primary branch flow paths 544 branches into a plurality of secondary branch flow paths 546. The liquid flow path system 540 is configured to transport a liquid (e.g., a liquid containing separated compounds) to each of the micropores 554 in the detection region 550.

[0054] As shown in FIG. 2 , the detection region 550 may include multiple sub-detection regions 552, such as a first sub-detection region 552A, a second sub-detection region 552B, a third sub-detection region 552C, and a fourth sub-detection region 552D. While four sub-detection regions are illustrated, the present disclosure may include more or fewer sub-detection regions. Each sub-detection region 552 includes multiple micropores 554. For example, the first sub-detection region 552A includes multiple first micropores 554A, the second sub-detection region 552B includes multiple second micropores 554B, the third sub-detection region 552C includes multiple third micropores 554C, and the fourth sub-detection region 552D includes multiple fourth micropores 554D. While each sub-detection region is illustrated as including four micropores, the sub-detection regions of the present disclosure may include more or fewer micropores.

[0055] In some embodiments, cells, such as an artificially cultured cell line, are added to the micropores 554 to detect whether the cells respond to the isolated compound. For example, cells of different cancer cell lines may be added to different micropores 554, and after incubation with the isolated compound for a certain period of time, e.g., several hours to several days, the activity and morphological changes of the cells in each micropore 554 may be observed using an optical instrument. In some embodiments, fluorescence in the micropores may be detected using a fluorescence microscope. In some embodiments, electrochemical signals in each micropore may be detected. In some embodiments, by controlling the reagent addition / sampling channel 260 and the third valve element 450, the liquid in these micropores 554 can be directed to an effluent collection module (722, see FIG. 12 ) outside the microfluidic chip 10, allowing secreted products from incubation of the cells with the isolated compound to be detected. In some embodiments, the value of a specific secreted product over time may be monitored using a liquid-phase chromatography device, a gas-phase chromatography device, a mass spectrometer, or the like. In some other embodiments, each micropore 554 may be provided with an antibody, a reactive color reagent, a signal sensing element, an analog, or a combination thereof to detect or select compounds with specific reactive properties.

[0056] 4 and 7, in several embodiments of the present disclosure, a heating and weighing component 110 is further provided below the process chamber 430 for heating and weighing the liquid in the process chamber 430. As shown in FIG. 4, the heating and weighing component 110 is located in the substrate layer 100 and includes a heating element 110A, a weighing element 110B, and a plurality of electrodes 112. In other embodiments, the heating and weighing component 110 may include either the heating element 110A or the weighing element 110B.

[0057] In some embodiments, heating element 110A is used to evaporate a portion of the extraction solvent to concentrate the sample extraction solution before introducing the extraction solution into the subsequent first filter element 510 and chromatography column 520. Upon heating, first valve element 420 may be opened, allowing the evaporated extraction solvent to exit microfluidic chip 10 through first valve element 420.

[0058] In some embodiments, the weighing element 110B includes a piezoelectric material and is used to sense the weight of the fluid in the processing chamber 430. The weighing element 110B is used to determine the weight of the extraction solution that has entered the subsequent chromatography column. Determining the weight of the extraction solution may be used to test and adjust the operating program conditions and compound separation effects of the microfluidic chip 10.

[0059] In some embodiments, the heating and weighing component 110 includes a heating element 110A and a weighing element 110B made of a piezoelectric material that is attached to the heating element 110A.

[0060] 1, 7, and 8A-8C, in various embodiments of the present disclosure, the flow of liquid in the microfluidic chip 10 is primarily driven via an actuator element 470 above the processing chamber 430, and the flow in the processing chamber 430 toward the sample injection channel 410 or the first filter element 510 may be controlled by adjusting the opening and closing of the first valve element 420 or the second valve element 440. When the actuator element 470 is raised, negative pressure is created in the processing chamber 430, allowing liquid to flow into the processing chamber 430 when the first valve element 420 is opened. When the actuator element 470 is depressed, positive pressure is created in the processing chamber 430, allowing liquid to flow out when the first valve element 420 or the second valve element 440 is opened.

[0061] In some embodiments, after the sample and extraction solution are mixed by suction and vortexing, the first valve element 420 is closed, the second valve element 440 is opened, and the liquid flows into the first filter element 510. Pressure is then applied, forcing the liquid into the chromatography column 520. The pressure is then reapplied, forcing the liquid into the second filter element 530. After the liquid flows out of the second filter element 530, the target compound can be selected based on the time difference between the flows. Optionally, the liquid is discharged via the reagent addition / sampling channel 260, or the third valve element 450 is closed and pressurized to enter the liquid flow system 540, and the compound is introduced into each micropore 554 in the detection region 550.

[0062] In several embodiments of the present disclosure, integrating the processing chamber 430 and the chromatography column 520 into the microfluidic chip 10 not only improves the integration of each functional component in the microfluidic chip 10, but also eliminates dead volume due to interfaces and avoids problems such as sample loss and contamination during offline processing. In some embodiments, sample concentration and weight determination may be performed in the processing chamber 430 prior to chromatography to improve the separation effect of the chromatography. The flow of liquid in the microfluidic chip 10 is controlled by the actuator element 470 and each valve element, and the elastic membrane can be controlled by air pressure to control the positive, negative, or normal pressure state of the liquid flow path to guide the liquid flow.

[0063] 3, the microfluidic chip 10 has a stack structure including a substrate layer 100, a first flow path layer 200 above the substrate layer 100, and a second flow path layer 300 above the first flow path layer 200. In some embodiments, the substrate layer 100 and the first flow path layer 200 may be connected by bonding. The first flow path layer 200 and the second flow path layer 300 may be connected by bonding.

[0064] In some embodiments, the material of the substrate layer 100 is a transparent, rigid material, such as glass, acrylic (PMMA), polycarbonate (PC), polystyrene (PS), engineering plastic (ABS), or other polymer plastics. In some embodiments, the thickness of the substrate layer 100 may be in the range of about 0.1 to about 1 millimeter (mm).

[0065] In some embodiments, the material of the first flow path layer 200 and the second flow path layer 300 is a transparent, soft material, such as polydimethylsiloxane (PDMS). In some embodiments, the thickness of the first flow path layer 200 may be in the range of about 0.1 to about 1 millimeter (mm), and the thickness of the second flow path layer 300 may be in the range of about 1 to about 10 millimeters (mm). In some embodiments, the thickness of the first flow path layer 200 may be approximately equal to the thickness of the second flow path layer 300, or the thickness of the first flow path layer may be less than the thickness of the second flow path layer 300.

[0066] In some embodiments, the sample injection channel 410 provided above the second flow path layer 300 may have a sample injection channel wall 412 with a height of 10 to 100 millimeters to accommodate the biological material to be extracted. During the extraction process, the driving membrane 280 repeatedly flows and mixes the extraction solution from the processing chamber 430 to the sample injection channel 410. Therefore, to accommodate the biological material and the extraction solution and prevent liquid overflow, the sample injection channel 410 has a large space, for example, the area of ​​the sample injection channel 410 is larger than the area of ​​the processing chamber 430 or the surface area of ​​the driving membrane 280. In some embodiments, the area of ​​the sample injection channel 410 is smaller than about 5% of the area of ​​the microfluidic chip 10 because it must have sufficient space for chromatography and analytes.

[0067] 1, 3, and 4, the substrate layer 100 is located at the bottom of the microfluidic chip 10. The substrate layer 100 may be an opening in the first flow path layer 200 or the bottom surface of a downward-facing groove.

[0068] As shown in FIG. 4, the substrate layer 100 is provided with a heated weighing component 110 and a plurality of electrodes 112 connected to the heated weighing component in order to evaporate some of the solvent and determine the weight of the extracted solution before performing chromatography.

[0069] 1, 3, and 5, in some embodiments, the first flow path layer 200 is defined with a plurality of grooves for liquid flow, and the positions of these grooves correspond to the liquid flow path system 540. As shown in FIG. 11, which is a cross-sectional view of the microfluidic chip 10 of FIG. 7 taken along line E-E', the grooves span four secondary branch channels 546, i.e., two first secondary branch channels 546A and two second secondary branch channels 546B. The grooves for liquid flow are recessed downward with their openings facing upward, and the bottom surface of the second flow path layer 300 corresponds to the upper surface of the liquid flow path system 540.

[0070] As shown in FIG. 11 , the first secondary branch channel 546A and the second secondary branch channel 546B have different widths. The transport path of the first secondary branch channel 546A is short and is used to transport liquid to the micropore 554A of the first sub-detection region 552A and the micropore 554B of the second sub-detection region 552B. The transport path of the second secondary branch channel 546B is long and is used to transport liquid to the micropore 554C of the third sub-detection region 552C and the micropore 554D of the fourth sub-detection region 552D. The width W1 of the first secondary branch channel 546A is larger than the width W2 of the second secondary branch channel 546B. In some embodiments, the size of the width W1 is, for example, about 1 millimeter to about 2 millimeters, and the size of the width W2 is, for example, 10 to 100 micrometers, for example, 20 micrometers. The larger width W1 reduces the resistance of the liquid transported through the first secondary branch channel 546A, allowing it to be transported quickly to the first and second detection sub-regions 552A, 552B, which are closer to the opening of the chromatography column 520. When all of the micropores 554 in the first and second detection sub-regions 552A, 552B are filled, the liquid begins to be transported through the second secondary branch channel 546B, which has a larger resistance, and then to the micropores 554 in the third and fourth detection sub-regions 552C, 552D. This configuration allows compounds eluted from the chromatography column 520 at different times to be introduced into the micropores 554 in different sub-detection regions 552, respectively.

[0071] The first flow path layer 200 further includes a first sample injection channel opening 210, a processing chamber groove 220, a first filter groove 230 for accommodating the first filter 514 of the first filter element 510, a second filter groove 232 for accommodating the second filter 534 of the second filter element 530, a reagent addition / sampling channel 260, a waste fluid channel 270, and a plurality of first micro through-holes 254 in the detection region 550. The first flow path layer 200 is further defined with an opening 262 communicating with the reagent addition / sampling channel 260. The first flow path layer 200 is further defined with an opening 272 communicating with the waste fluid channel 270.

[0072] The first flow path layer 200 further includes a plurality of hollowed-out regions, each of which has a thin film on top and a groove below the thin film. The material of the first flow path layer 200 is PDMS, and the thin film is deformable and elastic, so that raising and lowering the thin film creates negative or positive pressure in the flow path or processing chamber below the thin film, thereby driving the flow of fluid through the channels or chambers of the microfluidic chip 10. In some embodiments, the thin film regions of the first flow path layer 200 include the driving membrane 280 of the actuator element 470 above the processing chamber 430 (i.e., the processing chamber groove 220) and the elastic membrane above each valve element. As shown in FIG. 5, the first flow path layer 200 has a first elastic membrane 282 at the location of the first valve element 420, a second elastic membrane 284 at the location of the second valve element 440, a third elastic membrane 286 at the location of the third valve element 450, and a fourth elastic membrane 288 at the location of the fourth valve element 460.

[0073] In some embodiments, the valve elements on both sides of the processing chamber include valve columns that completely seal off the flow of liquid when the valve elements are closed. For example, when a vortex is generated by the actuator element 470 to separate the sample and extraction solvent, the second valve element 440 must be closed to prevent air from being drawn into the processing chamber 430 from the first filter element 510, creating bubbles. Referring to FIGS. 8A-8C , the first flow path layer 200 defines a first valve column 290 and a first elastic membrane 282 that is higher than the first valve column 290 of the first valve element 420. The first flow path layer 200 also defines a second valve column 292 and a second elastic membrane 284 that is higher than the second valve column 292 of the second valve element 440. The bottom surfaces of the first valve column 290 and the second valve column 292 are not connected to the substrate layer 100. Deformation of the elastic membrane may move the valve column into or out of contact with the substrate layer 100, thus opening or closing the valve element.

[0074] In some embodiments, the first flow path layer defines the third elastic membrane 286 of the third valve element 450 and the fourth elastic membrane 288 of the fourth valve element 460, and does not include a valve column. When the third elastic membrane 286 of the third valve element 450 is not deformed, the lower reagent addition / sampling flow path 260 remains open; when it is necessary to close the reagent addition / sampling flow path 260, positive pressure is applied to press down on the third elastic membrane 286 to close the flow path. Similarly, when the fourth elastic membrane 288 of the fourth valve element 460 is not deformed, the lower waste flow path 270 remains open; when it is necessary to close the waste flow path 270, positive pressure is applied to press down on the fourth elastic membrane 288 to close the flow path.

[0075] 9A and 9B illustrate a drive membrane 280 according to some embodiments. FIG. 9A is a bottom view of the drive membrane 280, and FIG. 9B is a cross-sectional view of the drive membrane 280 along line F-F'. The bottom surface 280S of the drive membrane 280 has an uneven structure with multiple patterns 610. In some embodiments, the pattern 610 may include multiple recessed portions, multiple protruding portions, or a combination thereof. The uneven bottom surface 280S of the drive membrane 280 is used to generate stronger turbulence to improve the mixing efficiency of the sample and extraction solvent when the drive membrane 280 is repeatedly deformed to mix the sample and extraction solvent. In some embodiments, the pattern 610 may include a recessed portion 612. In some embodiments, the depth of the recessed portion may be approximately one-third to two-thirds the thickness of the drive membrane 280. The recessed portion 612 shown in FIG. 9A is annular. 9C , the lower surface 280S of the drive membrane 280 has multiple patterns, where pattern 610A is a triangular pattern with recessed portions 612A, and pattern 610B is a rectangular pattern with recessed portions 612B. In other embodiments, the lower surface 280S of the drive membrane 280 may have other shaped patterns, such as a polygonal pattern (e.g., square, pentagonal, hexagonal, octagonal, or the like), an elliptical pattern, an irregular pattern, or a combination thereof. In other embodiments, the lower surface 280S of the drive membrane 280 may include more or fewer recessed portions 612.

[0076] 1, 3, and 6, in some embodiments, the second flow path layer 300 is defined with a plurality of grooves and vent holes for gas flow, the positions of these grooves corresponding to the gas flow paths, and the vent holes are positions where the microfluidic chip operating system 700 (see FIG. 12) introduces gas into the microfluidic chip 10. In some embodiments, as shown in FIG. 10, which is a cross-sectional view of the microfluidic chip 10 of FIG. 7 taken along line D-D', the plurality of grooves for gas flow are recessed upward with their openings facing downward, and the top surface of the first flow path layer 200 is the lower surface of the gas flow paths. In some embodiments, the second flow path layer 300 is further defined with an inlet / outlet port 362 and a waste outlet 372 corresponding to the openings 262 and 272 in the first flow path layer 200, respectively.

[0077] 7 and the cross-sectional views of FIGS. 8A to 8C, in some embodiments, the recessed groove for gas flow includes a drive control groove 320 above the drive membrane 280 on the processing chamber 430, and a drive gas flow path 322 communicating with the drive control groove 320. The second flow path layer 300 further includes a drive gas vent 324 communicating with the drive gas flow path 322.

[0078] In some embodiments, the gas flow grooves further include valve control grooves above the valves and elastic membranes, and valve gas flow channels connected to the valve control grooves. The second flow channel layer 300 further includes a plurality of valve gas vent holes each connected to a plurality of valve gas flow channels.

[0079] As shown in FIGS. 6 and 7, in the first valve element 420, a corresponding first valve control groove 340A is defined in the second flow path layer 300. The second flow path layer 300 also defines a first valve gas flow path 342A that communicates with the first valve control groove 340A, and a first valve vent 344A that communicates with the first valve gas flow path 342A. In the second valve element 440, a corresponding second valve control groove 340B is defined in the second flow path layer 300. The second flow path layer 300 also defines a second valve gas flow path 342B that communicates with the second valve control groove 340B, and a second valve vent 344B that communicates with the second valve gas flow path 342B. In the third valve element 450, a corresponding third valve control groove 340C is defined in the second flow path layer 300. Additionally, a third valve gas flow path 342C communicating with the third valve control groove 340C, and a third valve vent 344C communicating with the third valve gas flow path 342C are defined in the second flow path layer 300. In the fourth valve element 460, a corresponding fourth valve control groove 340D is defined in the second flow path layer 300. Additionally, a fourth valve gas flow path 342D communicating with the fourth valve control groove 340D, and a fourth valve vent 344D communicating with the fourth valve gas flow path 342D are defined in the second flow path layer 300.

[0080] The second flow path layer 300 further includes a second sample injection groove opening 310, a third filter groove 330 for accommodating the first filter 514 of the first filter element 510, a fourth filter groove 332 for accommodating the second filter 534 of the second filter element 530, and a plurality of second micro through-holes 354 in the detection region 550.

[0081] The first filter element 510 and the second filter element 530 may be provided at both ends of the chromatography column 520, respectively, and used to filter impurities in a liquid. For example, the first filter element 510 can prevent impurities from entering the chromatography column 520. The second filter element 530 can prevent filler material (e.g., beads) of the chromatography column 520 from flowing out to the detection region 550. In some embodiments, the material of the first filter 514 of the first filter element 510 and the second filter 534 of the second filter element 530 may be, for example, filter paper, cotton, synthetic fiber, a microchannel structure, or the like. In some embodiments, the pore size of the first filter element 510 and the second filter element 530 may be, for example, 0.01 to 10 micrometers.

[0082] The second flow path layer 300 further includes a chromatography column 520 containing a packing material for adsorbing specific compounds in the extraction solution. In some embodiments, the packing material can be, for example, silica gel, alumina, activated carbon, polyamide, macroporous adsorption resin, cross-linked glucose gel, agarose gel, polyacrylamide gel, polystyrene gel, metal-organic framework material, or the like.

[0083] In some embodiments, the first flow path layer 200 and the second flow path layer 300 may be formed using three-dimensional mold transfer to create openings, grooves, and cutouts in the first flow path layer 200 and the second flow path layer 300.

[0084] In some embodiments, after the openings and recessed grooves in the second flow path layer 300 are formed, the packing material is packed into the chromatography column 520 in the second flow path layer 300 .

[0085] In some embodiments, the substrate layer 100 and the first flow path layer 200 are bonded together by electropulp bonding. Then, the first filter 514 and the second filter 534 are provided in the first filter groove 230 and the second filter groove 232 of the first flow path layer 200, respectively. Then, the first flow path layer 200 and the second flow path layer 300 are bonded together by electropulp bonding.

[0086] Please refer to FIGS. 3 and 7, which show the structural arrangement of each layer in each region after the substrate layer 100, the first flow path layer 200, and the second flow path layer 300 are assembled.

[0087] The positions of the first sample injection channel opening 210 of the first flow path layer 200 and the second sample injection channel opening 310 of the second flow path layer 300 correspond to the sample injection channel 410, and the top surface of the substrate layer 100 is the bottom surface of the sample injection channel 410. The sample injection channel 410 further includes a sample injection channel wall 412 that is higher than the second flow path layer 300.

[0088] The positions of the valves and elastic membranes on the first flow path layer 200 and the positions of the valve control grooves on the second flow path layer 300 correspond to the valve elements.

[0089] The positions of the processing chamber groove 220 and the drive film 280 in the first flow path layer 200 and the position of the drive control groove 320 in the second flow path layer 300 correspond to the positions of the processing chambers 430 .

[0090] The positions of the plurality of first micro through-holes 254 in the first flow path layer 200 and the positions of the plurality of second micro through-holes 354 in the second flow path layer 300 correspond to the plurality of micro holes 554 in the detection region 550, respectively.

[0091] 8A-8C are cross-sectional views of the connecting lines AB and BC in FIG. 7, showing the area including the sample injection channel 410, the first valve element 420, the processing chamber 430, and the second valve element 440, and are schematic diagrams of the liquid 20 under different air pressure control conditions.

[0092] 8A, under normal pressure, the first elastic membrane 282 of the first valve element 420 is in an equilibrium position, and the first valve column 290 and the second valve column 292 are in a closed state. Also, the actuator element 470 (i.e., the drive membrane 280) is in an equilibrium position under normal pressure, and the drive membrane 280 is in a flat state.

[0093] As shown in Figure 8B, when the valve drive module (712 in Figure 12) of the microfluidic chip operating system applies a vacuum suction force to the first valve control groove 340A, the first valve control groove 340A becomes negative pressure, which further drives the first elastic membrane 282 to rise, thereby separating the first valve column 290 from the substrate layer 100 and causing the liquid 20 in the sample injection groove 410 to flow into the processing chamber 430. In addition, when the actuator element drive module (710 in Figure 12) also applies a vacuum suction force to the drive control groove 320, the drive control groove 320 becomes negative pressure, which further drives the drive membrane 280 to rise.

[0094] As shown in Figure 8C, when the actuator element driving module (712 in Figure 12) of the microfluidic chip operating system applies positive air pressure to the driving control groove 320, the driving control groove 320 becomes positive pressure, further pressing down the driving membrane 280 and causing the liquid 20 to flow through the first valve column 290 to the sample injection groove 410.

[0095] 12 illustrates a microfluidic chip operating system 700 according to some embodiments of the present disclosure. The microfluidic chip operating system 700 includes a microfluidic chip 10, a control module 702, an actuator element driving module 710, a valve driving module 712, a sensing module 714, a reagent addition module 720, an effluent collection module 722, a waste channel 724, and a detection device 730.

[0096] The control module 702 can control the operation of each module on the microfluidic chip 10 and receive the sensing signal of the microfluidic chip 10 sensed by the sensing module 714 .

[0097] The actuator element driving module 710 is connected to the driving gas vent 324 of the microfluidic chip 10 and may provide positive atmospheric pressure, negative atmospheric pressure or normal pressure to the driving control groove 320 to control the raising and lowering of the actuator element 470 (i.e., the driving membrane 280) above the processing chamber 430 of the microfluidic chip 10.

[0098] The valve driving module 712 is connected to each valve vent hole of the microfluidic chip 10, and may provide positive atmospheric pressure, negative atmospheric pressure, or normal pressure to each valve control groove so as to adjust the elevation of the elastic membrane of each valve element of the microfluidic chip 10, thereby controlling the opening and closing of the valve element.

[0099] The sensing module 714 is electrically connected to the micro-fluidic chip 10 and receives the sensing signals of the micro-fluidic chip 10 detected by the sensors, such as the temperature, pressure, or weight measured by the heating and weighing component 110 .

[0100] The discharged liquid collection module 722 is connected to the inlet / outlet 362 of the microfluidic chip 10 and is used to collect the liquid discharged from the microfluidic chip 10 .

[0101] The detection device 730 is in communication with the effluent collection module 722 .

[0102] The reagent addition module 720 is connected to the inlet / outlet 362 of the microfluidic chip 10 and transports the reagent to be added to the liquid flow channel system 540 and the detection region 550 of the microfluidic chip 10 .

[0103] FIG. 13 is a flowchart of a method of using the microfluidic chip 10 according to some embodiments.

[0104] In method 800, step 802 involves adding a sample tissue and an extraction solvent. In some embodiments, the sample tissue and extraction solvent are placed in the sample injection channel 410. For example, in plant sample tissue, there is a certain affinity between natural compound components and tissue cells. To dissolve the target component, the extraction solvent must have a greater affinity for the target component, eliminating the affinity adsorption between the target component and tissue cells and transferring the target component to the extraction solvent. In some embodiments, to improve extraction efficiency, the plant sample may first be dried (e.g., air-dried) or crushed. In some embodiments, to improve the extraction rate of the target component, an appropriate amount of acid, alkali, surfactant, etc. may be added to the solvent to aid desorption. After desorption, the chemical components disperse in the extraction solvent in the form of ions, molecules, etc., and soluble components dissolve in the extraction solvent according to their solubility. In some embodiments, the extraction solvent may be water, a hydrophilic organic solvent, a lipophilic organic solvent, etc., depending on the solubility of the target component in the extraction solvent. In some embodiments, the extraction solvent may be, for example, water, methanol, ethanol, acetone, n-butanol, ethyl acetate, ether, chloroform, dichloroethane, benzene, carbon tetrachloride, petroleum ether, or the like, or a combination thereof. In some embodiments, a co-solvent is further added to increase the solubility of the target component, remove certain specific impurities, improve component stability, etc. In some embodiments, the co-solvent may be, for example, an acid (e.g., hydrochloric acid, sulfuric acid, glacial acetic acid, tartaric acid, etc.), an alkali (ammonia water, carbonic acid, etc.), or a surfactant (e.g., Tween-20, Tween 80, etc.).

[0105] In step 804, cells are added to the plurality of micropores 554 of the detection region 550. In some embodiments, the cells may be cultured in the micropores 554 in the detection region of the microfluidic chip. In some embodiments, the cells are added to the plurality of micropores of the detection region 550 at least several hours (e.g., 4 hours) before extraction, allowing the cells to adhere to the bottom surface of the micropores.

[0106] In step 806, the first valve element 420 of the microfluidic chip 10 is opened, and the actuator element 470 is actuated to perform a vortex mixing process in the processing chamber 430. In some embodiments, the second valve element is closed, the first valve element is opened, and the actuator element's actuation membrane is raised, increasing the volume of the processing chamber and reducing the pressure, allowing fluid in the sample injection channel to flow into the processing chamber. Due to the height limitation of the opening of the first valve column of the first valve element, tissue fragments cannot enter the processing chamber. In some embodiments, repeated suction and vortex mixing increases the penetration of the extraction solvent into the sample tissue and the dissolution of components. In some embodiments, the suction and mixing time may be, for example, 5 to 30 minutes, e.g., about 20 minutes.

[0107] In step 808, a portion of the extraction solvent is heated and evaporated in the processing chamber 430 to concentrate the extraction solution.

[0108] In step 810, the heated weighing component 110 of the processing chamber 430 is used to weigh to determine the weight of the extraction solution for subsequent chromatography.

[0109] In step 812 , the second valve element 440 is opened, allowing the extraction solution to enter the first filter element 510 .

[0110] In some embodiments, the method further includes adding water or a buffer solution, or a solvent in which the target component is soluble, to the sample injection channel 410 before opening the second valve element 440 to ensure that a sufficient amount of liquid enters the downstream chromatography column 520.

[0111] In step 814 , actuator element 470 is controlled to apply pressure, forcing the extraction solution into chromatography column 520 .

[0112] In step 816 , actuator element 470 is controlled to re-pressurize, causing the effluent of chromatography column 520 to enter second filter element 530 .

[0113] The target compounds are separated by time difference in step 818. At the predetermined collection time, the third valve element 450 is closed, allowing the extract to flow into the liquid flow path system 540.

[0114] In step 820, repressurization causes the extracted effluent to flow in the liquid flow path system 540.

[0115] In step 822 , the extracted effluent containing the compounds is introduced into a plurality of micropores 554 containing the cells in the detection region 550 .

[0116] In step 824, the compound is co-incubated with the cells for a period of time, after which the cellular response is detected.

[0117] One embodiment of the present disclosure provides a method for operating a microfluidic chip, the method including: adding a sample and an extraction solvent to a sample injection channel of the microfluidic chip; adding a plurality of cells to each of a plurality of microholes in a detection region of the microfluidic chip; driving an actuator element of the microfluidic chip to mix the sample and the extraction solvent to form an extraction solution; transporting the extraction solution to a chromatography column in the microfluidic chip; transporting at least one target compound in the extraction solution that has flowed out of the chromatography column to the microholes; and detecting a response of the cells to the at least one target compound.

[0118] In some embodiments, the method of using the microfluidic chip further comprises evaporating a portion of the extraction solvent by a heater of the microfluidic chip before transporting the extraction solution to a chromatography column in the microfluidic chip.

[0119] In some embodiments, the method of using the microfluidic chip further comprises measuring the weight of the extraction solution with a sensor of the microfluidic chip before transporting the extraction solution to a chromatography column in the microfluidic chip.

[0120] In some embodiments, detecting the response of the cells to the at least one compound of interest comprises detecting secretions of the cells at the plurality of micropores with a detection instrument.

[0121] The present disclosure is disclosed in the embodiments as described above, but the above-described embodiments are not used to limit the present disclosure, and any person skilled in the art can make various changes and modifications without departing from the spirit and scope of the present disclosure, and the scope of protection of the present disclosure should be based on what is defined by the scope of the patent application attached later. [Explanation of symbols]

[0122] 10: Microfluidic chip 100: Substrate layer 110: Heating and weighing component 110A: Heating element 110B: weighing element 112: Electrode 20:Liquid 200: First flow path layer 210: First sample injection channel opening 220: Processing chamber groove 230: First filter groove 232: Second filter groove 254: First micro through hole 260: Reagent addition / sampling channel 270: Waste liquid flow path 262:Aperture 272:Aperture 280: Driving membrane 280S: Bottom surface 282: First elastic membrane 284: Second elastic membrane 286: Third elastic membrane 288: Fourth Elastic Membrane 290: First valve column 292: Second valve column 300: Second flow path layer 310: Second sample injection channel opening 320: Drive control groove 322: Driving gas flow path 324: Drive gas vent 330: Third filter groove 332: 4th filter groove 340A, 340B, 340C, 340D: Valve control groove 342A, 342B, 342C, 342D: Valve gas flow path 344A, 344B, 344C, 344D: Valve vents 354: Second micro through hole 362:Inflow / Outflow 372: Waste liquid outlet 410: Sample injection groove 412: Sample injection groove wall 420: First valve element 430: Processing room 440: Second valve element 450: 3rd valve element 460: 4th valve element 470: Actuator element 510: First filter element 512: First filtration chamber 514: First filter 520: Chromatography column 530: Second filter element 532: Second filtration chamber 534: Second filter 540: Liquid flow path system 542: Main channel 544: Primary branch flow path 546:Secondary branch flow path 546A: 1st secondary branch flow path 546B:Second secondary branch flow path 550: Detection area 552, 552A, 552B, 552C, 552D: Sub-detection area 554, 554A, 554B, 554C, 554D: Micro-hole 610, 610A, 610B: Pattern 612, 612A, 612B: recessed part 700: Microfluidic Chip Operating System 702: Control module 710: Actuator element drive module 712: Valve drive module 714: Sensing module 720: Reagent Addition Module 722: Effluent collection module 724: Waste fluid groove 730:Detection equipment 800: Method 802, 804, 806, 808, 810, 812, 814, 816, 818, 820, 822, 824: Process AB: line BC: Line D-D': Line E-E': Line F-F': line OD1: Opening dimension W1:Width W2:Width

Claims

1. a sample injection channel, a processing chamber, a first filter element, a chromatography column, a liquid flow system, and a detection region, which are connected in sequence; a first valve element interposed between the sample injection channel and the processing chamber; a second valve element interposed between the processing chamber and the first filter element; an actuator element disposed above the processing chamber, the actuator element including a driving membrane, configured to generate a vortex flow within the processing chamber and control a pressure within the processing chamber; A microfluidic chip comprising:

2. The microfluidic chip of claim 1 , wherein the first valve element comprises a first valve column having an opening dimension in the range of about 5 micrometers to about 30 micrometers.

3. The microfluidic chip according to claim 1 , wherein the lower surface of the driving membrane has an uneven structure.

4. The microfluidic chip of claim 1 , further comprising a heating element disposed below the processing chamber.

5. The microfluidic chip according to claim 1 , further comprising a weighing element provided below the processing chamber.

6. The microfluidic chip of claim 1 , further comprising a second filter element interposed between the chromatography column and the liquid flow path system.

7. a reagent addition / sampling channel communicating with the chromatography column; a third valve element configured to control the liquid flow direction and opening / closing of the reagent addition / sampling channel; The microfluidic chip of claim 1 , further comprising:

8. a waste liquid flow path communicating with the chromatography column; a fourth valve element configured to control opening and closing of the waste flow path; The microfluidic chip of claim 1 , further comprising:

9. The microfluidic chip comprises: a substrate layer; a first flow path layer above the substrate layer, the first flow path layer having a plurality of grooves defined therein for liquid flow; a second flow path layer above the first flow path layer, the second flow path layer having a plurality of grooves defined therein for gas flow; The microfluidic chip according to claim 1 , which has a stack structure comprising:

10. 10. The microfluidic chip of claim 9, further comprising a heated weighing component disposed within the substrate layer such that the heated weighing component is at the bottom of the process chamber.

11. 10. The microfluidic chip according to claim 9, wherein the actuator element further includes a drive control groove above the drive film, the processing chamber and the drive film being provided in the first flow path layer, and the drive control groove being provided in the second flow path layer.

12. The first valve element is a first valve column disposed between the sample injection channel and the processing chamber and provided on the first flow path layer; a first elastic membrane located higher than the first valve column and disposed on the first flow path layer; a first valve control groove located above the first valve column and the first elastic membrane and provided in the second flow path layer; The microfluidic chip of claim 9 .

13. The microfluidic chip according to claim 9 , wherein the chromatography column is provided in the second flow path layer.

14. The microfluidic chip according to claim 1 , wherein the volume of the sample injection channel is larger than the volume of the processing chamber.

15. The microfluidic chip according to any one of claims 1 to 14, an actuator element driving module configured to control the elevation and lowering of the actuator elements of the microfluidic chip; a valve driver module configured to control the opening and closing of the first valve element and the second valve element; A microfluidic chip operating system including:

16. The microfluidic chip operating system of claim 15 further comprising a detection device for detecting fluid from the chromatography column or the detection region.

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