WO2017107025A1

WO2017107025A1 - Detection device

Info

Publication number: WO2017107025A1
Application number: PCT/CN2015/098146
Authority: WO
Inventors: 黄荣堂
Original assignee: 黄荣堂
Priority date: 2015-12-21
Filing date: 2015-12-21
Publication date: 2017-06-29
Also published as: CN108474802A

Abstract

Disclosed is a detection device (10, 20, 30), comprising a base plate (100, 700), a cover body (200, 800), a micro-sensing chip (300, 900) and a micro-channel structure (400), wherein at least one of the base plate (100, 700) and the cover body (200, 800) contains a porous material. The base plate (100, 700) has a first surface (101), a groove (102, 702) is provided on the first surface (101), and the groove (102, 702) comprises a bottom part (103) and a slope (104), wherein the bottom part (103) is embedded into the base plate (100, 700), the slope (104) connects the first surface (101) with the bottom part (103) and is configured to one end, close to an injection port (402, 810), of the groove (102, 702). The cover body (200, 800) has a second surface (201) facing the first surface (101). The micro-sensing chip (300, 900) is embedded into the base plate (100, 700). The micro-channel structure (400) is embedded between the second surface (201) and the first surface (101) to form a micro-channel (401, 802). An object to be detected (600) enters from the injection port (402, 810) into the groove (102, 702) via the micro-channel (401, 802), and is separated into a lower layer liquid (601) and an upper layer liquid (602), wherein the lower layer liquid (601) is retained on the bottom part (103), and the upper layer liquid (602) flows from the groove (102, 702) to the micro-sensing chip (300, 900), and no additional power needs to be applied to the detection device (10, 20, 30) during the detection process of the object to be detected (600).

Description

Testing device

Technical field

The invention relates to a detecting device, in particular to a portable device and the test process is carried out without additional force, and the heavier molecules in the sample are separated from the lighter molecules and the related tests are simultaneously carried out. Detection device.

Background technique

In the conventional microfluidic detecting device, the sample can flow in the microchannel by the capillary phenomenon formed by the difference between the adhesion between the sample and the microchannel and the cohesive force of the sample itself. After the sample passes through a test zone in the microchannel, the micro-sensing chip in the test zone can be detected for the sample. Finally, the researchers transmitted the detection signals read by the micro-sensing chip to an external analytical instrument for related research and data analysis. However, today's detection devices must be powered to drive the flow of microfluids in the microchannels, thus greatly reducing the portability of the detection device.

In the detection procedure of some specific specimens, the researchers need to first screen the specimen to obtain specific molecules in the specimen, and then analyze and study the specific molecule. For example, when the sample is blood, in order to separate the blood cell from the plasma, the conventional detection method utilizes characteristics in which the blood cell and the plasma have different qualities, and the blood is separated into blood cells and plasma by a centrifugal separator. Therefore, the above-mentioned conventional technique is not only complicated, but also has a long detection time, and it is more necessary to provide the centrifugal separator power for detection, which is inconvenient.

Summary of the invention

In view of the above, an object of the present invention is to provide a detecting device capable of directly separating a sample into a mass-heavy molecule and a light-weight molecule, and detecting the sample of the separated molecule, so that centrifugation is not required The separator separates the sample, which not only simplifies the convenience of the detection procedure, but also has the concept of green energy saving.

Another object of the present invention is to provide a detecting device which is highly portable and does not require additional power during the detection of the sample.

In order to achieve the above object, the detecting apparatus of the present invention comprises: a substrate having a first surface, the first surface having a recessed portion, the recessed portion including a bottom portion and a slope, the bottom portion being embedded in the substrate, and the slope is connected to the first surface and a bottom portion and disposed at one end of the recess; a cover having a second surface facing the first surface; a micro-sensing chip embedded in the substrate; and a micro-channel structure embedded in the second surface, wherein the first surface and the second surface are mutually dense after the cover covers the substrate In combination, the microchannel structure is coupled to the first surface to form a microchannel comprising at least one injection port and a volume control slot for controlling the flow rate of the sample in the microchannel, the specimen entering the recess through the microchannel from the inlet The sample is separated into a lower layer liquid and an upper layer liquid in the depressed portion, the lower layer liquid stays at the bottom portion, and the upper layer liquid flows from the concave portion to the micro sensing chip; wherein the micro flow channel includes a first-class choke channel and a reaction tank The reaction tank is connected to the micro-sensing chip, and the flow-stop flow path is formed between the reaction tank and the capacity control tank; wherein at least one of the substrate and the cover body comprises a porous material.

In an embodiment of the invention, the sample is blood, the lower layer is blood cells, and the upper layer is plasma.

In an embodiment of the invention, one end of the micro flow channel has a capacity control slot for controlling the flow rate of the sample in the micro flow channel.

In an embodiment of the invention, the slope is disposed in an end of the recess adjacent to the injection port.

In an embodiment of the invention, the micro-sensing chip has at least one detecting structure for detecting bio-particles or bio-polymers in the structure to energize the sample.

In an embodiment of the invention, the detecting device further includes a plurality of terminals disposed on the substrate and connected to the micro sensing chip, and the plurality of terminals are coupled to a reading device.

In an embodiment of the invention, the plurality of terminals are connected to the micro-sensing chip by wire bonding.

In an embodiment of the invention, the detecting structure is a resistive type, a capacitive type, an impedance type, or a transistor type, or an electrochemical type, a counting type, or a photoelectric type based on a nano sensing material. The sensor, the nanomaterial is functionalized by a biopolymer selected from the group consisting of an antibody, an aptamer or a sugar molecule or an enzyme. In addition to being based on nano-sensing materials, the detection structure can also be selected from purely electrochemical or optoelectronic sensors.

In an embodiment of the invention, the nano-sensing material is selected from the group consisting of carbon nanotubes, graphene, reduced graphene oxide (rGO), graphene oxide (graphene oxide, GO), nanoribbon graphene, nano silicon wire, nano InP wire, nano GaN wire, nano semiconductor wire or nano semiconductor film.

In an embodiment of the invention, the material of the substrate is polymethylmethacrylate (PMMA), polyethylene terephthalate (PET), polycarbonate (PC), and porosity. Polydimethylsiloxane Polydimethylsilicon (PDMS), porous silica gel, rubber, plastic or glass.

In an embodiment of the invention, the cover body is made of polymethylmethacrylate (PMMA), porous polyethylene terephthalate (PET), polycarbonate (polycarbonate, PC). ), porous polydimethylsilicon (PDMS), porous silica gel, rubber or plastic.

In an embodiment of the invention, a pretreatment portion is provided between the recess and the injection port, and is adapted to separate the sample or mix with other reagents.

In an embodiment of the invention, when the second surface is assembled with the first surface, the edge of the second surface is located inside the side line of the first surface.

In an embodiment of the invention, the detecting means after the completion of the vacuuming process is packaged in a vacuum packaging bag.

In an embodiment of the invention, the first surface and the top surface of the micro-sense chip are in the same plane.

In an embodiment of the invention, the micro flow channel is located in a region between the micro sensing chip and the recess, and the channel cross-sectional area in the micro channel is reduced.

In order to achieve the above object, a detecting device of the present invention comprises: a substrate having a first surface comprising a recess and at least one reaction tank; a cover having a second surface covering the first surface of the cover; a micro-sensing chip embedded in the substrate, comprising at least one sensing region; a first injection port; a second injection port; and a micro-channel system formed between the substrate and the cover body, comprising a first micro-flow channel formed on a first surface of the substrate and connecting the reaction grooves and the second injection port, a second micro flow channel formed on the second surface of the cover body, and a capacity control groove formed on the second surface of the cover body and communicating with the second micro flow And a third micro flow channel connecting the reaction grooves to at least one sensing area of the micro sensing chip; wherein the second micro flow channel comprises a first portion and a second portion, the first portion is the first portion The injection inlet extends to the capacity control tank and communicates with the recess, and the second portion extends from the capacity control slot to the at least one reaction tank; wherein at least one of the substrate and the cover body comprises a porous material.

In order to achieve the above object, a detecting apparatus of the present invention includes: a substrate having a first surface including a recessed portion and a disintegrating groove; a cover having a second surface covering the first surface of the cover; The micro-sensing chip is embedded in the substrate and includes a sensing region; a micro-channel is formed between the substrate and the cover, and communicates with the sensing portion of the recess and the micro-sensing chip; and a heating component is formed under a portion of the micro-channel At least one of the substrate and the cover body comprises a porous material; wherein a decomposition liquid containing a sample having DNA fills the decomposition groove and the depressed portion, and then flows through the micro flow channel Above the hot component, the heated component is heated by circulation and copied in large quantities, and then flows through the microchannel to the sensing area of the micro sensing chip.

The above and other objects, features, and advantages of the present invention will be apparent from

DRAWINGS

1 is a perspective view of a detecting device according to an embodiment of the present invention.

2 is a perspective view of a substrate of a detecting device according to an embodiment of the present invention.

3 is a perspective view of a recessed portion of an embodiment of the present invention.

4 is a perspective exploded view of a substrate and a cover of a detecting device according to an embodiment of the invention.

FIG. 5 is a perspective exploded view of a substrate and a cover of a detecting device according to another embodiment of the present invention.

Fig. 6 is a schematic view showing a sample in a depressed portion according to an embodiment of the present invention.

FIG. 7 is a schematic diagram showing the reduction of the cross-sectional area in the microchannel according to an embodiment of the present invention;

FIG. 8 is a schematic diagram of a pre-processing unit according to an embodiment of the present invention.

Fig. 9 is a table showing an experimental record of a detecting device according to an embodiment of the present invention.

Figure 10 is a perspective view of another embodiment of the present invention.

11A and 11B are schematic top views of another embodiment of the present invention.

12 to 16 are different embodiments showing a depressed portion of a substrate according to another embodiment of the present invention.

Figure 17 is a top perspective view showing another embodiment of the present invention.

Figure 18 is a top perspective view showing another embodiment of the detecting device of the present invention.

Symbol Description

10, 20, 30: detection device

100, 700: substrate

101: first surface

102, 702: depression

103: bottom

104: Slope

200, 800: cover

300, 900: micro-sensing chip

814: first sensing cavity

816: second sensing cavity

818: third sensing cavity

820: fourth sensing cavity

400: Microchannel structure

401, 802, 804, 704: micro flow channel

402: injection port

403, 803: quantitative control structure

404: Pre-Processing Department

405, 805: capacity control slot

408, 808: flow resistance runner

409: Reaction tank

500, 720: terminal carrier

501, 721: a plurality of terminals

600: sample (blood)

601: lower layer (blood cell)

602: upper layer (plasma)

603: Biomarkers

604: aptamer

706: first reaction tank

708: second reaction tank

710: third reaction tank

712: fourth reaction tank

750: heating component

751: Heating chip

752: Resistance wire

752a, 752b: conductive end

760: Decomposition slot

809: Sensing cavity

810: first injection port

812: second injection port

detailed description

All technical and scientific terms used herein have the meaning as commonly understood by one of ordinary skill in the art, unless otherwise indicated.

The term "a" as used herein, unless otherwise specified, refers to an amount of at least one (one or more).

1 is a perspective view of a detecting device according to an embodiment of the present invention, FIG. 2 is a perspective view of a substrate of a detecting device according to an embodiment of the present invention, and FIG. 3 is a perspective view of a recessed portion according to an embodiment of the invention. FIG. 4 is a perspective exploded view of the substrate and the cover of the detecting device according to an embodiment of the present invention, and FIG. 5 is a perspective exploded view of the substrate and the cover of the detecting device according to another embodiment of the present invention. Referring to FIG. 1 , FIG. 2 , FIG. 3 , FIG. 4 and FIG. 5 , the detecting device 10 of the present invention includes a substrate 100 , a cover 200 , a micro sensing chip 300 , and a micro flow channel structure 400 . On the substrate 100, there is a first surface 101, and on the first surface 101, there is a recess 102. The recess 102 is composed of a bottom portion 103 and a slope 104, wherein the bottom portion 103 is embedded in the substrate 101, and the slope 104 is located at one end of the recess portion 102 near the injection port 402.

Further, in the embodiment of the present invention, at least one of the substrate 100 and the cover 200 comprises a porous material, and the cover 200 has a second surface 201 thereon. When the cover 200 covers the substrate 100, the first surface The 101 faces the second surface 201 and is in close contact with each other. The micro-channel structure 400 is disposed on the second surface 201. When the cover 200 covers the substrate 100, the micro-channel structure 400 is combined with the first surface 101 to form a micro-channel 401, at least the substrate 100 and the cover 200. After one of the porous material forms a vacuum state with the inside of the microchannel 401, the sample 600 is placed in the injection port 402 of the microchannel 401, and the sample 600 is driven from the injection by the suction generated by the vacuum in the microchannel 401. The inlet 402 enters the recess 102 via the microchannel 401, and the sample 600 is separated into the lower layer 601 and the upper layer 602 (shown in FIG. 6) in the recess 102. The lower layer 601 stays at the bottom 103, and the upper layer 602 The recessed portion 102 flows from the micro-sense chip 300.

In the embodiment of the present invention, the sample 600 may be a body fluid, including blood, cerebrospinal fluid, gastric juice and various digestive juices, semen, saliva, tears, sweat, urine, vaginal secretions, etc. or It is a solution containing the sample 600. In the present embodiment, blood is taken as an example. When blood enters the depressed portion 102 along the microchannel 401, the heavier blood cell (the lower layer 601) precipitates at the bottom 103 of the depressed portion 102, and the lighter plasma ( The supernatant liquid 602) will exit from the recess 102 and enter the micro-sensor chip 300 along the micro-channel 401. When the sample 600 is relatively thick, for example, blood, in order to make the sample 600 more smoothly injected from the injection port 402, an anticoagulant may be applied to the bottom and/or the side wall of the injection port 402.

It is worth mentioning that the slope 104 of the recess 102 of the present invention ensures that the specimen 600 can be concave. The trap 102 performs the function of separating. In the conventional technique, the sample 600 tends to have a misjudgment result when the separated sample 600 is interpreted by the micro-sensing chip 300 because the separation effect is not good. In the detecting device 10 of the present invention, the slope 104 of the depressed portion 102 near one end of the injection port 402 has a function of smoothing the flow of the sample 600 in the micro flow path 401 when the sample 600 is in the micro flow path 401. When flowing smoothly to the recess 102, molecules of different weights can produce different precipitation rates depending on the weight with relatively little external interference, and therefore, molecules of different weights can be separated more efficiently. In order to increase the effect of the recessed portion 102 separating the sample 600, an array of micropiles may be formed at an interface between the recessed portion 102 and the microchannel 401 at a distance of less than 3 micrometers to intercept a suspension of 3 micrometers or more. Or forming microcylinders spaced between 10 and 100 micrometers apart, and then complexing a plurality of microspheres having a size larger than the spacing of the plurality of microcylinders to form a plurality of voids to intercept suspended matter larger than the void size, or may increase The slope and/or surface roughness of the slope 104 of the recess 102 intercepts more suspended matter. The surfaces of the bottom portion 103 and the slope 104 of the depressed portion 102 may be treated with oxygen plasma or an intervening agent or the like to increase hydrophilicity and increase the probability of sedimentation of the suspended matter in the sample 600. If the sample 600 is blood containing platelets, a blood coagulant such as calcium chloride (CaCl) may be applied to the bottom portion 103 and the slope 104 of the depressed portion 102 to promote coagulation and aggregation of the blood cells to settle in the depressed portion 102. If the sample 600 is a non-blood sample such as a somatic cell in raw milk, it may be applied to the bottom portion 103 and the slope 104 of the depressed portion 102 or a mixture containing the thrombin (Thrombin) and fibrinogen may be added to the sample 600. (fibrinogen) and calcium ions (Calcium ion) to form a fibrin mesh, increasing the probability that the suspended matter settles in the depressed portion 102.

When the sample 600 is in the process of separation, the research project usually also includes quantitative analysis of the separated sample. In an embodiment of the invention, a volume control slot 405 is provided within the microchannel 401 relative to the other end of the injection port 402 for the purpose of analyzing the designed structure for the quantitative sample 600. When the sample 600 enters the microchannel 401 from the injection port 402, it passes through the recess 102 and the micro-sensing chip 300, and finally the sample 600 is stored in the capacity control slot 405. After the sample 600 is filled with the capacity control slot 405, the sample 600 at the injection port 402 does not enter the microchannel 401 again, so the signal detected by the micro-sensing chip 300 is controlled by the capacity control slot 405. The signal generated by the quantitative sample 600. In the present embodiment, it is assumed that the capacity control slot 405 has a capacity of 0.5 cc. Although the sample 600 applied to the injection port 402 is much larger than 0.5 cc, the signal sample 600 that can be detected by the micro-sense chip 300 is only 0.5. Cc. If the signal detected by the micro-sensing chip 300 is divided by 0.5 cc, the unit of the signal is presented in a concentration manner. In this embodiment, there is a certain amount of control structure 403 on the second surface 201, and the capacity control can be formed by the combination of the first surface 101 and the quantitative control structure 403. Slot 405.

In an embodiment of the invention, the micro-sensing chip 300 is embedded in the substrate 100, and the top surface of the micro-sensing chip 300 and the first surface 101 must be in the same plane to ensure that the sample 600 in the micro-channel 401 can be Flowing into the micro-sensing chip 300. In this embodiment, the micro flow channel 401 passes through at least one detecting structure of the micro sensing chip 300 from above, and in another embodiment, the micro flow channel 401 may also pass through at least one detecting structure of the micro sensing chip 300 from below. . Each detection structure can be quantified by bio-coupling modification, or can be further quantified by bio-particles or bio-polymers in the sample 600, or further via the micro-sensing chip 300, for example, a resistive type, a capacitive type, an impedance type, or a transistor type. Or electrochemical type comprising nano or non-nano, or counting type, photoelectric type comprising nano or non-nano sensing elements, converted into electrical signals, and finally the I/O pads of the micro sensing chip 300 are electrically connected to the plurality of terminals 501. The plurality of terminals 501 are electrically connected to the external reading device, and the detection signals are output to provide related research and analysis. In the embodiment of the present invention, the plurality of terminals 501 may also be connected to the micro-sensing chip 300 by wire bonding. In some embodiments, the micro-sensing chip 300 can also include an amplifier circuit to amplify the weak electronic signals detected.

When the sample 600 is measured using the detecting device 10, in order to perform the detection of the low-density sample 600, the present invention designs the micro-channel 401 in a region where the micro-channel 401 is located between the micro-sensing chip 300 and the depressed portion 102. The cross-sectional area of the inside is reduced, so that the flow rate of the sample 600 entering the micro-sensing chip 300 is lowered, so that the time during which the sample 600 stays in the micro-sensing chip 300 can be increased, and the majority of the sample 600 can be brought closer to the micro-sensing chip. 300, in order to facilitate the detection of low concentration sample 600. As shown in FIG. 7 , in an embodiment of the present invention, the flow path depth of the micro flow channel 401 is originally 60 μm, and the flow path depth between the micro sensing chip 300 and the recess portion 102 can be gently reduced by a slope. Up to 10 μm, so that the biomarker 603 of the deep flow channel (60 μm) upstream of the microchannel 401 can thus pass through the slope, slow down its flow rate, and limit its suspension range, and thus rush to the aptamer 604 at the bottom of the microchannel 401, Most of the biomarkers 603 are captured by the aptamer 604 at the bottom of the microchannel 401. Since the flow rate is low, the captured biomarkers 603 are fixed to the aptamer 604.

In the detecting device 10 of the present invention, the detecting structure is a resistive type, a capacitive type, an impedance type, or a transistor type, an electrochemical type, a counting type sensor based on a nano sensing material, and a nano sensing material. The biopolymer, in particular, refers to at least one of an antibody, or an aptamer, or a sugar molecule, or an enzyme molecule. The sensors can be of a plurality or array type to provide a quantitative test of a plurality of objects within the sample 600. The nano sensing material described above may be a material having semiconductor characteristics such as nanowires such as carbon nanotubes, nano silicon wires, nano InP wires, nano GaN wires, or nano semiconductor wires, which are suitable for sensing, Or a nano-semiconductor film, or graphene, reduced graphene oxide (rGO), graphene oxide (GO), nanoribbon graphene, and the like. In addition to being based on nano-sensing materials, the detection structure can also be selected from purely electrochemical or optoelectronic sensors.

In the embodiment of the detecting device 10 of the present invention, the material of the substrate 100 may be polymethylmethacrylate (PMMA), polyethylene terephthalate (PET), polycarbonate (PC). Porous polydimethylsilicon (PDMS), porous silica gel, rubber, plastic or glass; the material of the cover 200 may be polymethylmethacrylate (PMMA), polyethylene terephthalate Polyethylene terephthalate (PET), polycarbonate (PC), porous polydimethylsilicon (PDMS), porous silica gel, rubber or plastic.

It is to be noted that in selecting the material of the substrate 100 and the cover 200, it is necessary to consider the material properties between the substrate 100 and the cover 200. When the cover 200 covers the substrate 100, the inside of the microchannel 401 must be evacuated to form a vacuum state to provide a suction force for driving the sample 600 to flow in the microchannel 401. Therefore, at least one of the substrate 100 and the cover 200 must be a porous material. Preferably, the material hardness characteristic between the substrate 100 and the cover 200 can be a hard and a soft one. Taking an embodiment of the present invention as an example, the substrate 100 is made of a plastic having a higher hardness, and the cover 200 is made of a porous polydimethylsilicon (PDMS) having a lower hardness and is more rigid. The microchannel structure 400 is formed on the low cover 200, so when the microchannel 401 is evacuated, the porous polydimethylsilicon (PDMS) having a lower hardness is attached to the hardness. On the high plastic substrate 100, the air in the pores of the PDMS is simultaneously removed to maintain the vacuum state in the microchannel 401. When the detecting device 10 is exposed to normal atmospheric pressure, the pore vacuum state of the porous polydimethylsilicon (PDMS) is gradually filled by the outside air, but as long as it is not balanced with the external pressure, The microchannel 401 is provided with a negative pressure to drive the sample 600 to flow.

In the above embodiment, the substrate 100 is made of a plastic material having a relatively high hardness, and the material of the cover 200 is a polydimethylsilicon (PDMS) having a low hardness and a porosity. However, the present invention is not limited thereto, and the substrate 100 may also be made of a material having a lower hardness, and the cover 200 is a material having a higher hardness. It is to be noted that in order to enable the present invention to be mass-produced, in addition to selecting the material of the substrate 100 and the cover 200, it is necessary to apply the technique of injection molding, and additionally to prevent the substrate 100 which has been combined and evacuated. Cover 200 The collision occurs during the transportation and transportation so that the substrate 100 and the cover 200 are not tightly adhered to maintain the vacuum state. The present invention is specifically designed to laminate the first surface 101 and the second surface 201 to each other, and the edge of the second surface 201 is Located inside the side of the first surface 101 (as shown in Figure 1). Since the first surface 101 and the second surface 201 are seam-finished when they are attached to each other, in the mass production, the automatic device only needs to seal the slit with a sealant, and the vacuum can be applied to the injection port to make the micro-injection A vacuum state is formed inside the flow path. In an embodiment of the invention, the detection device 10 after completion of the vacuuming process is packaged in a vacuum package.

In another embodiment of the present invention, the recess 102, the micro-sense chip 300, the micro-channel structure 400, and the quantitative control structure 403 are disposed on the substrate 100. In still another embodiment, the recess 102, the micro-feel The test chip 300, the micro flow channel structure 400, and the quantitative control structure 403 may also be disposed on the cover 200. In another preferred embodiment of the present invention, a plurality of microchannels 401, a plurality of recesses 102, a plurality of micro-sensing chips 300, and a plurality of quantitative control structures 403 may be disposed on the substrate 100 via the special substrate 100. In combination with the cover 200 configuration, the single detecting device 10 can simultaneously process a plurality of test samples, which is not only efficient but also saves time and cost; and in another preferred embodiment of the present invention, between the recess 102 and the injection port 402 There is a pre-processing portion 404 (shown in Figure 8) that is adapted to separate the sample 600 or to mix with other reagents. Some special specimens 600 must be removed from the original specimen 600 prior to actual detection, or mixed with other materials, and the prior treatment portion 404 of the present invention has the function of providing the above separation and mixing.

It should be noted that the detecting device 10 of the present invention utilizes a vacuuming method, and one of the substrate 100 and the cover 200 is made of a porous material, and the micro flow channel 401, the recess portion 102 and the capacity control slot 405 are formed inside. A negative pressure is generated with respect to the external atmospheric pressure. When the sample 600 is placed at the injection port 402, the sample 600 can flow through the concave portion 102 along the micro flow channel 401 by the pressure difference between the atmospheric pressure and the negative pressure. Sensing chip 300 to capacity control slot 405 completes this detection and does not require additional power to drive sample 600 to flow during the detection process using a power component such as a pump or valve member. 9 is an experimental recording table of the detecting device 10 according to an embodiment of the present invention. Referring to FIG. 9, in the test sample 1, since the detecting device 10 is not subjected to the vacuuming process first, the sample 600 is dropped to the injection port 402. After one day, the sample 600 still cannot flow to the depressed portion 102; in the test samples 2 to 6, after the vacuuming treatment of the detecting device 10 for 6 minutes and 30 seconds to 7 minutes, the sample 600 takes about 14 to 17 minutes. Flows to the entrance of the micro-sensing chip 300. Therefore, it has been experimentally found that after the vacuuming process of the detecting device 10 of the present invention is performed for a certain period of time, the flow pattern of the sample 600 in the detecting device 10 has consistency and repeatability.

Referring to FIG. 10, in another embodiment, a plurality of terminals of the detecting device 10 of the present invention may be formed. On a terminal carrier 500, the terminal carrier 500 is formed on the first surface 101 of the substrate 100 and protrudes from the side of the substrate 100. The micro-sensing chip 300 of the present embodiment can be similar to the foregoing embodiment; the cover 200, the capacity control slot 405, the injection port 402, the recess 102, and the micro flow channel 401 can also be similar to the foregoing embodiments.

Referring to FIG. 11A and FIG. 11B, the detecting apparatus 10 of the foregoing embodiments of the present invention may further include: a first-class choke 408 is formed in the micro-channel 401; and a reaction tank 409 is connected to the micro-sensing chip 300, wherein the flow resistance is The flow path 408 is formed between the capacity control tank 405 and the reaction tank 409 to further delay the time during which the sample 600 flows to the quantitative control structure 403, and increase the reaction time of the sample 600 and the micro-sensing chip 300, thereby improving the accuracy of the detection. . Since the sample 600 is filled with the sample liquid and the object to be tested is suspended in the sample liquid, it is necessary to have sufficient analyte to be precipitated on the micro-sensing chip 300 to make the most accurate analysis, and the flow resistance channel 408 can be used. After the sample 600 is filled in the reaction tank 409, the sample 600 is allowed to stay in the reaction tank 409 for 3 to 7 minutes, and in one embodiment, up to about 5 minutes. The effect of the flow resistance flow path 408 slowing down the flow rate of the sample 600 allows a certain amount of the sample 600 to fully react with the micro-sensing chip 300. When the cavity corresponding to the reaction tank 409 is filled with the sample 600, the sample 600 is The flow restricting flow path 408 continues to flow to the capacity control tank 405, but the volume of the flow restricting flow path 408 is very small relative to the reaction tank 40, so the sample 600 in the reaction tank 409 before the sample 600 flows to the capacity control tank 405 It can be regarded as a state of being stationary, that is, the micro-sensing chip 300 is reacted with a certain amount of the sample 600 in a unit time, and therefore the volume of the cavity in which the reaction tank 409 is combined with the substrate 100 can be used as a unit of a certain amount of analysis. The flow resistance flow path 408 may have a meandering pattern, as shown in FIG. 11A, the number of times of meandering is short, the meandering amplitude is short, and the width of the flow path is narrow, or the number of times of zigzag in FIG. 11B is small, the meandering amplitude is long, and the width of the flow path is wide. .

Referring to FIGS. 12 to 16, there are shown variations in the recess 102 of the substrate 100 of the detecting device 10 of the present invention. As shown in FIG. 12, the flat bottom 103 of the recess 102 is closer to the injection port 401 than the slope 104. As shown in FIG. 13, the recess 102 may have only the ramp 104 without a flat bottom, and the depth of the ramp 104 is deeper the further away from the injection port 401. As shown in FIG. 14, the recess 102 may have only the ramp 104 without a flat bottom, and the depth of the ramp 104 is shallower as it is farther from the injection port 401. As shown in FIG. 15, the width of the recessed portion 102 may be narrower as it is farther from the injection port 401. As shown in FIG. 16, the width of the recessed portion 102 may be wider as it is farther from the injection port 401.

In summary, the detecting device 10 of the present invention can separate the heavier molecules and the lighter molecules in the sample 600 by the design of the recess 102. Since it is not necessary to use a centrifugal separator, and the sample 600 can be directly separated, the detecting device 10 of the present invention has a green and environmentally friendly concept of convenience and energy saving. When the detecting device 10 of the present invention is electrically connected to the external device In the device, the separated sample 600 can upload the detection signal to the external device while the detection is performed, so that the researcher can perform subsequent related research and analysis, so the detection device 10 of the present invention also has the advantages of rapid detection and simple operation. The advantages.

Referring to the top perspective view of Fig. 17, another embodiment of the detecting device of the present invention is shown. The detecting device 20 of the present invention comprises: a substrate 700 comprising a recess 702, a microchannel 704 and at least one reaction tank, for example, comprising a first reaction tank 706, a second reaction tank 708, a third reaction tank 710 and A fourth reaction tank 712, wherein the first to fourth reaction tanks are connected by the micro flow passage 704; a cover 900 includes a plurality of injection ports such as a first injection port 810, a second injection port 812, and a micro flow channel 802. 804, a first sensing cavity 814, the at least one sensing cavity can include a second sensing cavity 816, a third sensing cavity 818, and a fourth sensing cavity 820, and a certain amount of control structure 803; The micro-sensing chip 900 is embedded in the substrate 700; and a terminal carrier 720 is connected to the micro-sensing chip 900 and a plurality of terminals 721 are formed on the surface. The microchannel 802 can include a first portion between the first injection port 810 and the quantitative control structure 803 and a second portion substantially parallel to the first portion and extending from the quantitative control structure 803 toward the second injection port 812, wherein the micro flow channel The first portion and the second portion of 802 are two different locations that are in communication with the quantitative control structure 803. The cover 800 can further include a first-stage choke 808 formed in the first portion of the micro flow channel 802. As can be seen from the previously described embodiments, the quantitative control structure 803 can be formed on the substrate 700 in addition to the cover 800.

The material of the substrate 700 can be similar to the plastic or hydrophilic material with higher hardness in the foregoing embodiment, and the material of the cover 800 can be similar to the porous PDMS or other hydrophobic material with soft hardness, that is, the cover. The body 800 is more hydrophobic than the substrate 700. After the cover 800 covers the substrate 700, the first portion of the micro flow channel 802 is a first sensing region connected to the recess 702 of the substrate 700 and the micro sensing chip 900, the first reaction tank 706 and the second reaction tank. 708, the third reaction tank 710 and the fourth reaction tank 712 are closed by the cover 800 and respectively communicated by the branches of the second portion of the micro flow passage 802, and the quantitative control structure 803 of the cover 800 forms a Capacity control slot 805. After the cover 800 covers the substrate 700, the first sensing cavity 814, the second sensing cavity 816, the third sensing cavity 818, and the fourth sensing cavity 820 are respectively covered in the second of the micro sensing chip 900. The four different sensing portions of the sensing region, the first sensing cavity 814, the second sensing cavity 816, the third sensing cavity 818, and the fourth sensing cavity 820 are respectively connected to each other through a micro flow channel 804 A reaction tank 706, a second reaction tank 708, a third reaction tank 710, and a fourth reaction tank 712. The first portion of the microfluidic channel 802 is selectively widened at the corresponding micro-sensing chip 900. The

microchannels

802, 804, 704 and the capacity control slot 805 can be considered as a microchannel system. Although the detecting device 20 of the embodiment has four reaction tanks and four sensing chambers, in other embodiments, the detecting device 20 may have only one counter. Should be a slot and a sensing cavity. Although the detecting device 20 of the embodiment has two sensing regions, in other embodiments, only one sensing region may be provided. Since the cover 800 has hydrophobicity and the substrate 700 has hydrophilicity, when a sample is introduced into the microchannel 802 from the first injection port 810, the sample is automatically taken by one side of the substrate 700 in the micro flow channel 802. The sample is adsorbed until the substrate 700 adsorbs the sample beyond its hydrophilic saturation value, and the sample fills the microchannel 802. Therefore, when the sample flows to a tank such as the recess 702 or the volume control slot 805, After filling the tank, the microchannel 802 flows out and flows to the next tank.

The detecting device 20 of the present embodiment can be applied to drug resistance detection, such as the following steps:

Step 1: The first reaction tank 706, the second reaction tank 708, the third reaction tank 710, and the fourth reaction tank 712 are prefilled with four antibiotics (not shown) for one bacteria, and are attached to the patch form. The bottom of each tank.

Step 2: The first injection port 810 is dropped into the sample (not shown), and the sample enters the depressed portion 702 through the first portion of the micro flow channel 802 to filter impurities. It can be seen from the foregoing embodiment that the vacuum state of the microchannel 802 of the detecting device 20 can automatically flow the sample in the direction of the capacity control slot 805. If necessary, the cover 800 can press the capacity control slot 805 to generate a deformation to form a negative pressure to drive the detection. The flow of the body.

Step 3: After the recessed portion 702 is filled, the filtered sample reaches the first sensing region of the micro-sensing chip 900 via the first portion of the micro-flow channel 802. At this time, the micro-sensing chip 900 first determines whether the sample is in the sample. Have bacteria to detect drug resistance.

Step 4: The culture solution is dropped from the second injection port 812. Since the first reaction tank 706, the second reaction tank 708, the third reaction tank 710, and the fourth reaction tank 712 are connected by the micro flow passage 704, the communication can be utilized. The tube principle controls the liquid level of the four reaction tanks to a height of 95% of the reaction tank.

Step 5: The filtered sample is injected into the capacity control slot 805 after being initially interpreted by the micro-sensing chip 900. After the volume control slot 805 collects the filtered sample, the filtered sample begins to pass through the microflow. The four branches of the second portion of the track 802 are injected into the first reaction tank 706, the second reaction tank 708, the third reaction tank 710, and the fourth reaction tank 71, respectively, and after the four reaction tanks are filled, the filtered The sample then fills the first sensing cavity 814, the second sensing cavity 816, the third sensing cavity 818, and the fourth sensing cavity 82 via the microchannel 804.

Step 6: Read four electrical signals of the four sensing portions of the second sensing area of the micro-sensing chip 900.

Step 7: After the bacteria in the first reaction tank 706, the second reaction tank 708, the third reaction tank 710, and the fourth reaction tank 71 are cultured for about half an hour, the fourth sensing region of the micro-sensing chip 900 is read again. The four electrical signals of the sensing department.

Step 8: Compare the electrical signals of Step 6 and Step 7. If there is a significant change, you can judge the corresponding Antibiotic resistance. For example, in Step 7, if the electrical signal corresponding to the first sensing cavity 814 of the micro-sensing chip 900 is increased compared to Step 6, it means that the bacteria are resistant to the antibiotics in the first reaction tank 706, and vice versa in Step 7 if the micro-sensing chip 900 corresponds. The electrical signal of the first sensing cavity 814 has no significant increase compared to Step 6, and the antibiotic in the first reaction tank 706 has an effect of inhibiting the growth of the bacteria.

The detecting device 20 of the present embodiment can also be applied to detecting exosomes in blood, as follows:

Step 1: The blood with the extracellular chromosome is dropped from the first injection port 810 (not shown). After the blood enters the depressed portion 702 through the first portion of the microchannel, the blood cell is intercepted by the depressed portion 702, and the plasma continues to proceed. The micro-sensing chip 900 flows. It can be seen from the foregoing embodiment that the vacuum state of the microchannel 802 of the detecting device 20 can automatically flow the plasma toward the volume control tank 805. If necessary, the lid 800 can press the capacity control tank 805 to form a negative pressure to drive the flow of plasma.

Step 2: The cell decomposition solution (1ysis buffer) is dropped from the second injection port 812, and the first reaction tank 706, the second reaction tank 708, the third reaction tank 710, and the fourth reaction tank 712 are subjected to the micro flow channel 704. Connected, based on the principle of the communication tube, the liquid level of the four reaction tanks is controlled to a height of 95% of the reaction tank. Since the detecting device 20 is applied to detect the foreign body in the blood, it is not necessary to determine whether the foreign body is a preset type. Therefore, the micro sensing chip 900 of the detecting device 20 may have only one sensing region, one reaction tank, and communication with the micro device. One of the sensing chips 900 senses the cavity.

Step 3: The volume control tank 805 collects the plasma from the first portion of the microchannel 802, and the plasma is injected into the first reaction tank 706 and the second reaction tank 708 from the four branches of the second portion of the microchannel 803, respectively. The third reaction tank 710 and the fourth reaction tank 712 react with the cell decomposition liquid, and the cell decomposition liquid can decompose the cell wall of the foreign body to expose the protein of the foreign body. After the four reaction tanks are filled, the reacted plasma then fills the first sensing chamber 814, the second sensing chamber 816, the third sensing chamber 818, and the fourth sensing chamber 82 via the microchannel 804. .

Step 4: Read four electrical signals of the four sensing portions of the second sensing region of the micro-sensing chip 900.

Step 5: After the external reaction in the four reaction tanks starts to react for about half an hour, the four electrical signals of the four sensing portions of the second sensing region of the micro-sensing chip 900 are read again.

Step 6: Compare the electrical signals of Step 5 and Step 4, and estimate the concentration of the foreign body from the change of the two-step electrical signal.

Referring to Figure 18, there is shown a top perspective view of another embodiment of the detecting device of the present invention. The detecting device 30 includes a substrate 700 including a recessed portion 702 and a decomposition groove 760; a cover 800 includes a micro flow channel 802, a sensing cavity 809, a first injection port 810 and a second injection port 812; A micro-sensing chip 900 is embedded in the substrate 700 and has a sensing region connected by the sensing cavity 809 A heating module 750 includes a heating chip 751 and a resistor wire 752 formed on the heating chip 751 in a meandering manner; and a terminal carrier 720 is connected to the micro sensing chip 900 and a plurality of terminals 721 are formed on the surface. The recessed portion 702 may be a recessed portion of the foregoing embodiments, such as different forms of FIGS. 12 to 16, and/or formed with arrays of micropillars (not shown) at the interface with the microfluidic channel 802. Or other disclosed structures may increase the sediment retention probability of the specimen. The electric resistance wire 752 of the heating assembly 750 has two

conductive ends

752a, 752b electrically connected to two contacts of a power supply (not shown), and the temperature of the electric resistance wire 752 rises to heat the upper micro flow channel 802 to heat. The chip 751 can regulate the temperature of the resistor line 752. The detecting device 30 may include a capacity control slot 805. The quantitative control structure may be formed on the cover 800 or the substrate 700. The first-class choke 808 is formed in the micro flow channel 802 and located in the capacity control slot 805 and the sensing cavity 809. between.

The detecting device 30 of the present embodiment can be applied to the detection of DNA extracted by Polymerase Chain Reaction (PCR), and a sample having DNA such as blood (not shown) can be injected into the decomposition tank 760 by the first injection port 810. A cell decomposing liquid can be injected from the second injection port 812. After the cell decomposition liquid fills the decomposition tank 760, the sample can flow from the micro flow channel 802 to the depressed portion 702, and the sample can be filtered in the concave portion 702, and then heated by the micro flow channel 802 to the heating assembly 750, and the micro flow The track 802 is meander-shaped on the heating assembly 750 and its meandering direction is substantially perpendicular to the meandering direction of the electric resistance wire 752, so that the sample can be uniformly heated. After the sample is heated by the heating element 750 for a plurality of cycles between 95 degrees C and 65 degrees C, the contained DNA is largely replicated, and then flows from the microchannel 802 to the sensing cavity 809 and with the micro-sensing chip 900. In response, the micro-sensing chip 900 may also have a plurality of sensing portions for different detection of DNA. As in the previous embodiment, the flow restricting flow path 808 is combined with the capacity control groove 805 to allow the detecting device 30 to quantitatively analyze the sample.

The present invention has been disclosed in the above embodiments, but it is not intended to limit the scope of the present invention, and the present invention may be modified and retouched without departing from the spirit of the invention. The scope of protection is subject to the definition of the appended claims.

Claims

A detecting device comprising:

a substrate having a first surface having a recessed portion, the recessed portion including a bottom portion and a slope, the bottom portion being embedded in the substrate, the slope connecting the first surface and the bottom portion and disposed on the substrate One end of the recess;

a cover having a second surface facing the first surface;

a micro-sensing chip embedded in the substrate;

a micro flow channel structure is embedded in the second surface, wherein after the cover body covers the substrate, the first surface and the second surface are in close contact with each other, so that the micro flow channel structure is coupled with the first surface The flow channel includes at least one injection port and a volume control groove for controlling a flow rate of the sample in the micro flow channel, the sample enters the concave portion from the injection port through the micro flow channel, and the sample body is separated in the concave portion Forming a lower layer liquid and an upper layer liquid, the lower layer liquid stays at the bottom portion, and the upper layer liquid flows from the depressed portion to the micro sensing chip;

Wherein the microchannel comprises a first-stage choke and a reaction tank, the reaction tank is connected to the micro-sensing chip, and the flow-stop flow path is formed between the reaction tank and the capacity control tank;

At least one of the substrate and the cover comprises a porous material.
The detecting device according to claim 1, wherein the sample is blood, the lower layer is blood cells, and the upper layer is plasma.
The detecting device according to claim 1, wherein the slope is disposed in an end of the recess adjacent to the injection port.
The detection device of claim 1 wherein the micro-sensing chip has at least one detection structure that energizes biological particles or biopolymers in the sample.
The detecting device of claim 1 further comprising a plurality of terminals disposed on the substrate and coupled to the micro-sensing chip, the terminals being coupled to a reading device.
The detecting device according to claim 5, wherein the terminals are connected to the micro sensing chip by wire bonding.
The detecting device according to claim 5, wherein the detecting structure is a resistive type, a capacitive type, or a transistor type or an electrochemical sensor based on a nano sensing material, and the nano material passes through a biopolymer Functionalized, the biopolymer is selected from an antibody, aptamer or sugar molecule or enzyme, or the detection structure comprises a non-nano electrochemical or optoelectronic sensor.
The detecting device according to claim 7, wherein the nano sensing material is selected from the group consisting of carbon nanotubes, graphene, reduced graphene oxide (rGO), graphene oxide (graphene oxide) , GO), nanoribbon graphene, nano silicon wire, nano InP wire, nano GaN wire, nano semiconductor wire or nano semiconductor film.
The detecting device according to claim 1, wherein the material of the substrate is polymethylmethacrylate (PMMA), polyethylene terephthalate (PET), polycarbonate (PC), poly. Polydimethylsilicon (PDMS), porous silica gel, rubber, plastic or glass.
The detecting device according to claim 1, wherein the cover is made of polymethylmethacrylate (PMMA), polyethylene terephthalate (PET), polycarbonate (PC), Polydimethylsilicon (PDMS), porous silica gel, rubber or plastic.
The detecting device according to claim 1, wherein a pretreatment portion is provided between the depressed portion and the injection port, and is adapted to separate the sample or mix with other reagents.
The detecting device according to claim 1, wherein the second surface and the first When the surface assembly is combined, the edge of the second surface is located inside the side line of the first surface.
The detecting device according to claim 1, wherein the bottom portion and the slope of the depressed portion are coated with calcium chloride or a mixture comprising thrombin, fibrinogen, and calcium ion to form a Fibrin mesh.
The detecting device according to claim 1, wherein the micro-cylinders arranged in an array in the intersection of the depressed portion and the micro-flow passage are spaced apart from each other by less than 3 μm, or a boundary between the depressed portion and the micro-flow passage is formed. The microcylinders arranged in an array are spaced apart from each other by 10 to 100 micrometers, and a plurality of microspheres having a size larger than the spacing between the microcylinders form a plurality of voids to intercept the suspended matter in the specimen.
The detecting device according to claim 1, wherein the microchannel is located in a region between the micro-sensing chip and the recess, and a channel cross-sectional area in the micro-channel is reduced.
The detecting device according to claim 1, wherein the slope is disposed in an end of the recess away from the injection port.
The detecting device according to claim 1, wherein the flow restricting flow path is in a meandering pattern.
The detecting device according to claim 1, wherein the width of the depressed portion is gradually widened away from the injection opening in a plan view or narrowed away from the injection opening.
A detecting device comprising:

a substrate having a first surface comprising a recess and at least one reaction tank;

a cover having a second surface covering the first surface of the cover;

a micro-sensing chip is embedded in the substrate, and includes at least one sensing region;

a first injection port;

a second injection port;

a micro flow channel system is formed between the substrate and the cover body, and includes a first micro flow channel formed on the first surface of the substrate and communicating the reaction grooves and the second injection port, a second micro flow channel is formed on the second surface of the cover body and a capacity control groove is formed on the second surface of the cover body and communicates with the second micro flow channel, and a third micro flow channel connects the Responding to the at least one sensing region of the micro sensing chip;

The second microchannel includes a first portion and a second portion. The first portion extends from the first injection port to the capacity control slot and communicates with the recess. The second portion is The capacity control tank extends to the at least one reaction tank; wherein at least one of the substrate and the cover comprises a porous material.
The detecting device according to claim 19, wherein the cover is more hydrophobic than the substrate.
The detecting device according to claim 19, wherein the microchannel system, the at least one reaction tank, and the recess are in a vacuum state before being detected.
The detecting device according to claim 19, wherein the cover body is adapted to be deformed at a position corresponding to the capacity control groove.
The detecting device according to claim 19, further comprising a first-stage choke formed in the first portion of the second microchannel.
The detecting device of claim 19, further comprising a terminal carrier electrically connected to the micro sensing chip and having a plurality of terminals.
The detecting device according to claim 19, wherein the sensing region comprises a first sensing region connected by the first portion of the second microchannel and a second sensing region by the third microchannel Connected.
A detecting device comprising:

a substrate having a first surface including a recess and a decomposition groove;

a cover having a second surface covering the first surface of the cover;

A micro-sensing chip is embedded in the substrate and includes a sensing area;

a micro flow channel is formed between the substrate and the cover body, and communicates the recessed portion and the sensing region of the micro-sensing chip;

a heating assembly formed below a portion of the microchannel;

One of the decomposing liquids containing a DNA-filled sample fills the decomposing tank and the recessed portion, and then flows through the micro-flow passage to the heating assembly, is heated by the heating assembly and is largely replicated, and then flows through the micro-flow passage. Going to the sensing area of the micro-sensing chip;

At least one of the substrate and the cover comprises a porous material.
The detecting device according to claim 26, wherein the micro-pillars arranged in an array in the boundary between the depressed portion and the micro-flow passage are spaced apart from each other by less than 3 μm, or a boundary between the depressed portion and the micro-flow passage is formed. The microcylinders arranged in an array are spaced apart from each other by 10 to 100 micrometers, and a plurality of microspheres having a size larger than the spacing between the microcylinders form a plurality of voids to intercept the suspended matter in the specimen.
The detecting device according to claim 26, further comprising a first injection port for injecting the sample and a second injection port to inject the decomposition liquid.
The detecting device of claim 26, further comprising a first-class choke, a capacity control slot and a sensing cavity, wherein the sensing cavity is connected to the microchannel and communicates with the sensing region, the flow obstruction A track is formed in the microchannel and between the capacity control slot and the sensing chamber.
The detecting device according to claim 26, wherein the heating assembly comprises a heating chip and a resistance wire located on the heating chip and having a meandering shape, the portion of the micro flow channel located on the resistance wire is also meandered and The meandering direction is perpendicular to the meandering direction of the resistance wire.