Microwell array chip, method of using same, and detection device

Abstract

A microwell array chip, a method for using the same, and a detection device are provided. The microwell array chip includes a microwell array substrate, the microwell array substrate including a first main surface and a second main surface provided opposite to each other, n reaction chambers, and an idle region, the n reaction chambers are arranged in an array in the microwell array substrate, the idle region is provided surrounding the n reaction chambers, the reaction chambers are configured to accommodate a sample to be measured, the shape of the orthogonal projection of the reaction chambers onto a first reference plane on which the first main surface is located is a regular N-gon, the area of ​​the idle region is divided into n' virtual units, and the shape of the orthogonal projection of the virtual units onto the first reference plane is the same as the shape of the orthogonal projection of the reaction chambers onto the first reference plane. The microwell array chip can efficiently utilize the area of ​​the microwell array substrate, thereby increasing the volume of a single reaction chamber, and further improving the sensitivity of the microwell array chip and reducing the detection limit of the microwell array chip.

Description

[Technical field]

[0001] SUMMARY OF THE DISCLOSURE Embodiments of the present disclosure relate to a microwell array chip, a method for using the same, and a detection device. [Background technology]

[0002] Polymerase chain reaction (PCR) is a molecular biology technique that can amplify DNA fragments in vitro. Digital PCR technology (dPCR) is a new technology for absolute quantification of nucleic acid molecules, which does not rely on standard curves or reference samples and can directly detect the copy number of target molecules without setting up controls. The principle of digital PCR technology is to distribute low template reagents into a large number of microwells, and after statistical analysis, it is found that most of the microwells have no target molecules or only one, and after the amplification is completed, the initial template amount is obtained by counting by the optical detection module. Compared with traditional quantitative PCR, digital PCR technology has higher sensitivity, specificity, high tolerance and accuracy, and this technology is widely used in the detection of ultra-trace nucleic acid samples, CNV analysis and gene expression detection of complex samples.

[0003] A microwell array chip is a substrate that contains an array formed of multiple microwells, where each microwell can act as a small test tube and can be used to detect or select a particular compound from many compounds. Thus, digital PCR technology can use microwell array chips to perform detection, by forcing reagents into the chip wells and distributing the reagents to a large number of microwells. Summary of the Invention [Means for solving the problem]

[0004] The embodiment of the present disclosure provides a microwell array chip, a method for using the same, and a detection device. The microwell array chip includes a microwell array substrate, the microwell array substrate includes a first main surface and a second main surface that are provided opposite to each other, n reaction chambers, and a virtual idle area, the n reaction chambers are arranged in an array in the microwell array substrate, the virtual idle area is provided surrounding the n reaction chambers, the reaction chambers are configured to accommodate a sample to be measured, the area of ​​the virtual idle area is divided into n' virtual units, the shape of the orthogonal projection onto a first reference plane on which the first main surface of the reaction chambers is located is a regular N-gon, and the shape of the orthogonal projection onto the first reference plane of the virtual units is the same as the shape of the orthogonal projection onto the first reference plane of the reaction chambers. The microwell array chip can increase the volume of a single reaction chamber by efficiently utilizing the area of ​​the microwell array substrate, and can further improve the sensitivity of the microwell array chip and reduce the detection limit of the microwell array chip.

[0005] At least one embodiment of the present disclosure provides a microwell array chip including: a microwell array substrate including a first and a second main surface arranged opposite each other; n reaction chambers arranged in an array within the microwell array substrate and configured to accommodate a sample to be measured, the n reaction chambers having a shape of a regular N-gon when orthogonally projected onto a first reference plane on which the first main surface is located; and a virtual idle region surrounding the n reaction chambers, the area of ​​the virtual idle region being divided into n' virtual units, the shape of the orthogonal projection of the virtual units onto the first reference plane being the same as the shape of the orthogonal projection of the reaction chambers onto the first reference plane, and a total volume V of the n reaction chambers satisfying Equation 1.

[0006]

number

[0007] For example, a microwell array chip according to one embodiment of the present disclosure further includes a support region configured to provide a support structure, and the total volume V of the n reaction chambers satisfies Equation 2.

[0008]

number

[0009] For example, the microwell array chip according to one embodiment of the present disclosure further includes a reaction region disposed around the support region, and the n reaction chambers are located in the reaction region.

[0010] For example, in a microwell array chip according to one embodiment of the present disclosure, in the reaction region, the center distances of the orthogonal projections of two adjacent reaction chambers onto the first reference plane are equal.

[0011] For example, in a microwell array chip according to one embodiment of the present disclosure, the shape of the orthogonal projection of the reaction chamber onto a first reference plane on which the first main surface is located and the shape of the orthogonal projection of the virtual unit onto the first reference plane are both regular hexagons, and the total volume V of the n reaction chambers satisfies Equation 3.

[0012]

number

[0013] For example, in a microwell array chip according to one embodiment of the present disclosure, the value of n ranges from 8,000 to 100,000.

[0014] For example, in a microwell array chip according to one embodiment of the present disclosure, the total volume V of the n reaction chambers satisfies Equation 4.

[0015]

number

[0016] For example, a microwell array chip according to one embodiment of the present disclosure further includes a first hydrophobic layer located on the first main surface, and an orthogonal projection of the first hydrophobic layer onto the first reference plane is spaced apart from an orthogonal projection of the reaction chamber onto the first reference plane.

[0017] For example, a microwell array chip according to one embodiment of the present disclosure further includes a second hydrophobic layer located on the second major surface and extending to an edge of the reaction chamber.

[0018] For example, a microwell array chip according to one embodiment of the present disclosure further includes a second hydrophobic layer, wherein the orthogonal projection of the second main surface of the second hydrophobic layer onto a second reference plane includes an opening, and an edge of the opening overlaps with an edge of the reaction chamber on the second reference plane.

[0019] For example, a microwell array chip according to one embodiment of the present disclosure further includes a second hydrophobic layer located on the second main surface, the reaction chamber penetrates the microwell array substrate in a direction perpendicular to the first reference plane, the second hydrophobic layer spans the reaction chamber, and the orthogonal projection of the reaction chamber onto the second reference plane on which the second main surface is located is within the orthogonal projection of the second hydrophobic layer onto the second reference plane.

[0020] For example, in a microwell array chip according to one embodiment of the present disclosure, the reaction chamber is concave from the first main surface into the microwell array substrate and has a body bottom located within the microwell array substrate, and the distance between the body bottom and the first reference plane is less than the thickness of the microwell array substrate.

[0021] For example, in a microwell array chip according to one embodiment of the present disclosure, the contact angle between the first hydrophobic layer and the sample to be measured is less than the critical angle of the reaction chamber, and the critical angle is the angle between an extension of the side wall of the reaction chamber and a tangent to the surface of the sample to be measured that contacts the side wall of the reaction chamber.

[0022] For example, in a microwell array chip according to one embodiment of the present disclosure, the bottom of the body includes at least one exhaust hole, each of which penetrates the bottom of the body in a direction perpendicular to the first reference plane.

[0023] For example, in a microwell array chip according to one embodiment of the present disclosure, the bottom of the body includes one exhaust hole, and the orthogonal projection of the exhaust hole onto a second reference plane on which the second main surface is located is located at the center of the orthogonal projection of the bottom of the body onto the second reference plane.

[0024] For example, in a microwell array chip according to one embodiment of the present disclosure, the bottom of the body includes a plurality of the exhaust holes, and the orthogonal projections of the exhaust holes onto a second reference plane in which the second main surfaces are located are arranged around the center of the orthogonal projection of the bottom of the body onto the second reference plane.

[0025] For example, a microwell array chip according to one embodiment of the present disclosure further includes a dialysis membrane and a second hydrophobic layer, wherein the reaction chamber penetrates the microwell array substrate in a direction perpendicular to the first reference plane, the second hydrophobic layer extends to the edge of the reaction chamber, and the dialysis membrane spans the reaction chamber.

[0026] For example, in a microwell array chip according to one embodiment of the present disclosure, the orthogonal projection onto a second reference plane on which the second main surface of the reaction chamber is located is within the orthogonal projection onto the second reference plane of the dialysis membrane.

[0027] For example, in a microwell array chip according to one embodiment of the present disclosure, the dialysis membrane is a flexible dialysis membrane.

[0028] For example, in a microwell array chip according to one embodiment of the present disclosure, the dialysis membrane is located on the side of the second hydrophobic layer adjacent to the second main surface.

[0029] For example, in a microwell array chip according to one embodiment of the present disclosure, the dialysis membrane is located on the side of the second hydrophobic layer away from the second main surface.

[0030] For example, in a microwell array chip according to one embodiment of the present disclosure, the angle between the inner side surface of the reaction chamber and the first main surface is greater than 90 degrees.

[0031] For example, in a microwell array chip according to one embodiment of the present disclosure, the inner surface of the reaction chamber includes a first sub-surface and a second sub-surface in a direction perpendicular to the first reference plane, the second sub-surface is located on the side of the first sub-surface away from the first main surface, the included angle between the first sub-surface and the first main surface is greater than 90 degrees, and the included angle between the second sub-surface and the second main surface is greater than 90 degrees.

[0032] For example, in a microwell array chip according to one embodiment of the present disclosure, the inner surface of the reaction chamber includes a first sub-surface, a second sub-surface, and a third sub-surface in a direction perpendicular to the first reference plane, the second sub-surface is located on the side of the first sub-surface away from the first main surface, the third sub-surface is located on the side of the second sub-surface away from the first sub-surface, the included angle between the first sub-surface and the first main surface is greater than 90 degrees, the plane on which the second sub-surface is located is perpendicular to the first reference plane, and the included angle between the third sub-surface and the second main surface is greater than 90 degrees.

[0033] For example, in a microwell array chip according to one embodiment of the present disclosure, the inner surface of the reaction chamber includes a first sub-surface, a second sub-surface, and a third sub-surface in a direction perpendicular to the first reference plane, the second sub-surface is located on the side of the first sub-surface away from the first main surface, the third sub-surface is located on the side of the second sub-surface away from the first sub-surface, the included angle between the first sub-surface and the first main surface is greater than 90 degrees, the second sub-surface is an arcuate surface concave toward the microwell array substrate, and the included angle between the third sub-surface and the second main surface is greater than 90 degrees.

[0034] For example, in a microwell array chip according to one embodiment of the present disclosure, a first hydrophilic film and a second hydrophilic film are provided on the inner surface of the reaction chamber, the first hydrophilic film and the second hydrophilic film are provided adjacent to each other in a direction perpendicular to the first reference plane, the surface of the first hydrophilic film away from the inner surface of the reaction chamber is an arcuate surface convex toward the central axis of the reaction chamber, and the surface of the second hydrophilic film away from the inner surface of the reaction chamber is an arcuate surface convex toward the central axis of the reaction chamber.

[0035] For example, in a microwell array chip according to one embodiment of the present disclosure, the inner surface of the reaction chamber is flat, the thickness of the first hydrophilic film in a direction perpendicular to the inner surface varies, such that the surface of the first hydrophilic film moving away from the inner surface of the reaction chamber is an arcuate surface, and the thickness of the second hydrophilic film in a direction perpendicular to the inner surface varies, such that the surface of the second hydrophilic film moving away from the inner surface of the reaction chamber is an arcuate surface.

[0036] For example, in a microwell array chip according to one embodiment of the present disclosure, the inner surface of the reaction chamber includes a first sub-surface and a second sub-surface in a direction perpendicular to the first reference plane, the second sub-surface is located on the side of the first sub-surface away from the first main surface, and the first sub-surface is convex toward the central axis of the reaction chamber, such that the surface of the first hydrophilic film away from the inner surface of the reaction chamber is an arcuate surface, and the second sub-surface is convex toward the central axis of the reaction chamber, such that the surface of the second hydrophilic film away from the inner surface of the reaction chamber is an arcuate surface.

[0037] For example, in a microwell array chip according to one embodiment of the present disclosure, the shape of the orthogonal projection of the reaction chamber onto the first reference plane is virtually one of a circle, a regular hexagon, and a regular octagon.

[0038] For example, in a microwell array chip according to one embodiment of the present disclosure, the shape of the orthogonal projection of the reaction chamber onto the first reference plane is virtually a triangle.

[0039] For example, a microwell array chip according to one embodiment of the present disclosure further includes a first packaging membrane located on the side of the first main surface away from the second main surface, and a second packaging membrane located on the side of the second main surface away from the first main surface, and the first packaging membrane and the second packaging membrane are attached onto the microwell array substrate by static electricity or colloids.

[0040] For example, a microwell array chip according to one embodiment of the present disclosure further includes a photocurable oil at an opening position adjacent to the first main surface of the reaction chamber, the photocurable oil including a boss structure that is in contact with the first main surface and is located on the side of the first hydrophobic layer adjacent to the central axis of the reaction chamber.

[0041] For example, in a microwell array chip according to one embodiment of the present disclosure, the microwell array substrate is a flexible substrate.

[0042] For example, in a microwell array chip according to one embodiment of the present disclosure, the microwell array substrate further includes a liquid introduction flow path, the n reaction chambers are connected to the liquid introduction flow path, and a unidirectional membrane is provided between each of the reaction chambers and the liquid introduction flow path.

[0043] For example, in a microwell array chip according to one embodiment of the present disclosure, the liquid introduction channel includes a main liquid introduction channel and n branch liquid introduction channels each communicating with the main liquid introduction channel, and the n branch liquid introduction channels are provided in one-to-one correspondence with the n reaction chambers.

[0044] For example, in a microwell array chip according to one embodiment of the present disclosure, the microwell array substrate further includes a liquid introduction channel including a plurality of sub-liquid introduction channels which are interconnected, the sub-liquid introduction channels having different heights, and each of the sub-liquid introduction channels communicating with a plurality of reaction chambers.

[0045] For example, in a microwell array chip according to one embodiment of the present disclosure, the heights of the multiple sub-liquid introduction channels decrease in order.

[0046] For example, in a microwell array chip according to one embodiment of the present disclosure, the heights of the multiple liquid introduction channels decrease successively from the middle to both sides.

[0047] For example, a microwell array chip according to one embodiment of the present disclosure further includes a first substrate located on one side of the microwell array substrate and spaced apart from the first main surface, and a second substrate located on a side of the microwell array substrate away from the first substrate, the second substrate including a heating electrode, and an orthogonal projection of the heating electrode onto the first reference plane overlaps with the orthogonal projection of at least a portion of the n reaction chambers onto the first reference plane.

[0048] For example, in a microwell array chip according to one embodiment of the present disclosure, the first substrate includes a first base substrate and a third hydrophobic layer located on the side of the first base substrate adjacent to the second substrate.

[0049] For example, in a microwell array chip according to one embodiment of the present disclosure, the second substrate includes a second base substrate, a control electrode located on the second base substrate, a first insulating layer located on the side of the control electrode facing away from the second base substrate, and a second insulating layer, the first insulating layer includes a connection hole exposing at least a portion of the control electrode, the heating electrode is located on the side of the first insulating layer facing away from the second base substrate and is connected to the control electrode via the connection hole, the second insulating layer is located on the side of the heating electrode facing away from the first insulating layer, and the microwell array substrate is located on the second insulating layer.

[0050] For example, a microwell array chip according to one embodiment of the present disclosure further includes a photosensitive sensor located on a side of the second substrate away from the first substrate, the photosensitive sensor being configured to detect light emitted by the reaction chambers in the microwell array substrate.

[0051] At least one embodiment of the present disclosure further provides a detection device including the microwell array chip described in any one of the above.

[0052] For example, a detection device according to one embodiment of the present disclosure further includes a first casing located on one side of the microwell array chip and spaced apart from the microwell array chip, and a second casing located on the side of the microwell array chip away from the first casing and spaced apart from the microwell array chip, wherein the distance between the microwell array chip and the second casing is greater than or equal to the thickness of the microwell array chip.

[0053] For example, in a detection device according to one embodiment of the present disclosure, the second casing includes a support structure, the support structure includes a first platform portion and a second platform portion, the height of the second platform portion is greater than the height of the first platform portion, the first platform portion is configured to contact a bottom surface of the microwell array chip, and the second platform portion is configured to contact a side surface of the microwell array chip.

[0054] For example, in a detection device according to one embodiment of the present disclosure, the shape of the orthogonal projection of the first platform portion onto the first main surface includes an arc triangle, the shape of the orthogonal projection of the second platform portion onto the first main surface includes a semicircle, the base of the arc triangle connected to the semicircle is a straight line, and the remaining two sides of the arc triangle are arcs.

[0055] For example, in a detection device according to one embodiment of the present disclosure, the second casing further includes a positioning circular base configured to be placed in contact with a side surface of the microwell array chip.

[0056] At least one embodiment of the present disclosure further provides a method of using a microwell array chip, the microwell array chip including a microwell array substrate, the method comprising the steps of injecting a sample to be measured into the microwell array substrate and packaging the sample to be measured in the microwell array substrate, the microwell array substrate including n reaction chambers and an idle area, the n reaction chambers are arranged in an array in the microwell array substrate and configured to accommodate the sample to be measured, a shape of an orthogonal projection of the reaction chambers onto a first reference plane on which the first main surface is located is a regular N-gon, the idle area is arranged surrounding the n reaction chambers, an area of ​​the idle area is divided into n′ virtual units, a shape of the orthogonal projection of the virtual units onto the first reference plane is the same as a shape of the orthogonal projection of the reaction chambers onto the first reference plane, and a total volume V of the n reaction chambers satisfies Equation 1.

[0057]

number

[0058] For example, in a method of using a microwell array chip according to one embodiment of the present disclosure, the step of packaging the sample to be measured in the microwell array substrate includes the steps of injecting the sample to be measured into the microwell array substrate, and then attaching a first packaging film to the side of the first main surface facing away from the second main surface by static electricity or colloids, and attaching a second packaging film to the side of the second main surface facing away from the first main surface.

[0059] For example, in a method of using a microwell array chip according to one embodiment of the present disclosure, the step of packaging the sample to be measured in the microwell array substrate includes the steps of: injecting the sample to be measured into the microwell array substrate, coating a photocurable oil at an opening position adjacent to the first main surface of the reaction chamber, and curing the photocurable oil using ultraviolet light.

[0060] For example, in a method of using a microwell array chip according to one embodiment of the present disclosure, the microwell array substrate includes a first sub-flexible microwell array substrate and a second sub-flexible microwell array substrate, a plurality of sample flow paths are included between the first sub-flexible microwell array substrate and the second sub-flexible microwell array substrate, and the step of injecting a sample to be measured into the microwell array substrate includes the step of injecting the sample to be measured into the plurality of sample flow paths.

[0061] For example, a method of using a microwell array chip according to an embodiment of the present disclosure further includes the step of injecting a sample to be measured into the plurality of sample flow paths using a vacuum pumping method.

[0062] For example, in a method of using a microwell array chip according to one embodiment of the present disclosure, the step of packaging the sample to be measured in the microwell array substrate further includes the step of using rollers to partition each of the sample flow paths to form a plurality of reaction chambers and sealing the plurality of reaction chambers.

[0063] In order to more clearly describe the technical solutions of the embodiments of the present disclosure, the drawings of the embodiments are briefly described below. It is obvious that the drawings described below only relate to some embodiments of the present disclosure, but are not intended to limit the present disclosure. [Brief description of the drawings]

[0064] [Figure 1]FIG. 1 is a schematic plan view of a microwell array chip according to one embodiment of the present disclosure. [Diagram 2] FIG. 2 is a schematic cross-sectional view of a microwell array chip according to one embodiment of the present disclosure. [Figure 3A] FIG. 3A is a schematic plan view of a reaction chamber of another microwell array chip according to one embodiment of the present disclosure. [Figure 3B] FIG. 3B is a schematic cross-sectional view of a reaction chamber of another microwell array chip according to an embodiment of the present disclosure. [Figure 4A] FIG. 4A is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure. [Figure 4B] FIG. 4B is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Diagram 5] FIG. 5 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Figure 6A] FIG. 6A is a schematic plan view of another microwell array chip according to one embodiment of the present disclosure. [Figure 6B] FIG. 6B is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Figure 7A] FIG. 7A is a schematic plan view of another microwell array chip according to one embodiment of the present disclosure. [Figure 7B] FIG. 7B is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Figure 8A] FIG. 8A is a schematic plan view of another microwell array chip according to one embodiment of the present disclosure. [Figure 8B] FIG. 8B is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Figure 9A] FIG. 9A is a schematic plan view of another microwell array chip according to one embodiment of the present disclosure. [Figure 9B] FIG. 9B is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Figure 10A]FIG. 10A is a schematic plan view of another microwell array chip according to one embodiment of the present disclosure. [Figure 10B] FIG. 10B is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Figure 11A] FIG. 11A is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure. [Figure 11B] FIG. 11B is a schematic perspective view of a reaction chamber of a microwell array chip according to one embodiment of the present disclosure. [Figure 11C] FIG. 11C is a schematic perspective view of a reaction chamber of another microwell array chip according to an embodiment of the present disclosure. [Figure 12A] FIG. 12A is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure. [Figure 12B] FIG. 12B is a schematic perspective view of a reaction chamber of a microwell array chip according to one embodiment of the present disclosure. [Figure 12C] FIG. 12C is a schematic perspective view of a reaction chamber of another microwell array chip according to an embodiment of the present disclosure. [Figure 13A] FIG. 13A is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure. [Figure 13B] FIG. 13B is a schematic perspective view of a reaction chamber of a microwell array chip according to one embodiment of the present disclosure. [Figure 13C] FIG. 13C is a schematic perspective view of a reaction chamber of another microwell array chip according to an embodiment of the present disclosure. [Figure 14A] FIG. 14A is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure. [Figure 14B] FIG. 14B is a schematic perspective view of a reaction chamber of a microwell array chip according to one embodiment of the present disclosure. [Figure 14C] FIG. 14C is a schematic perspective view of a reaction chamber of another microwell array chip according to an embodiment of the present disclosure. [Figure 15]FIG. 15 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Figure 16] FIG. 16 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Figure 17] FIG. 17 is a schematic diagram of the structure of a microwell array chip according to one embodiment of the present disclosure. [Figure 18] FIG. 18 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Figure 19] FIG. 19 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Figure 20] FIG. 20 is a schematic plan view of another microwell array chip according to an embodiment of the present disclosure. [Figure 21] FIG. 21 is a schematic plan view of another microwell array chip according to an embodiment of the present disclosure. [Figure 22] FIG. 22 is a schematic plan view of another microwell array chip according to an embodiment of the present disclosure. [Figure 23] FIG. 23 is a schematic plan view of another microwell array chip according to an embodiment of the present disclosure. [Figure 24] FIG. 24 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Diagram 25] FIG. 25 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Figure 26] FIG. 26 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. [Figure 27] FIG. 27 is a schematic diagram of a detection device according to one embodiment of the present disclosure. [Figure 28] FIG. 28 is a schematic diagram of another detection device according to an embodiment of the present disclosure. [Figure 29] FIG. 29 is a schematic plan view of a second casing of a detection device according to an embodiment of the present disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0065] In order to make the objectives, technical solutions and advantages of the embodiments of the present disclosure clearer, the technical solutions of the embodiments of the present disclosure will be described clearly and completely below with reference to the drawings of the embodiments of the present disclosure. It is obvious that the described embodiments are only some embodiments of the present disclosure, but not all of the embodiments. Based on the described embodiments of the present disclosure, other embodiments that a person skilled in the art can obtain without making creative efforts all belong to the protection scope of the present disclosure.

[0066] Unless otherwise defined, technical or scientific terms used in this disclosure should have the ordinary meanings understood by those skilled in the art. The words "first", "second" and similar words used in this disclosure do not denote any order, number or importance, but are used only to distinguish different components. Similar words such as "comprise" or "comprises" mean that the element or object before the word covers the element or object listed after the word and its equivalents, and does not exclude other elements or objects. Similar words such as "connected" or "coupled" are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect.

[0067] Unless otherwise defined, the features such as "parallel", "vertical" and "same" used in the embodiments of the present disclosure include cases such as "parallel", "vertical", "same" in the strict sense, and cases including a certain error such as "approximately parallel", "approximately vertical", and "approximately the same". For example, the above "approximately" can indicate that the difference between the compared objects is within 10% or 5% of the average value of the compared objects. In the following description of the embodiments of the present disclosure, when the number of one member or element is not specifically indicated, it means that the member or element may be one, may be multiple, or can be understood as at least one. "At least one" means one or multiple, and "multiple" means at least two. "Provided in the same layer" in the embodiments of the present disclosure refers to the relationship between multiple film layers formed after the same material undergoes the same step (e.g., one-step patterning process). Here, "same layer" does not always mean that the thickness of multiple film layers is the same, or that the height of multiple film layers in a cross-sectional view is the same.

[0068] Through research, the inventors of the present application found that the main problem of the current digital PCR chip is how to improve the sensitivity and how to reduce the detection limit of the chip. From a chemical point of view, if the volume of a single chamber is large enough, the reaction is complete as long as the sample is sufficient, while if the volume of a single reaction chamber is large, it will cause a waste of chamber volume per unit area of ​​the chip. Therefore, it is necessary to control the volume of the reaction chamber within a certain range.

[0069] In view of this situation, the embodiments of the present disclosure provide a microwell array chip, a method for using the same, and a detection device, the microwell array chip comprising a microwell array substrate, the microwell array substrate comprising a first main surface and a second main surface arranged opposite to each other, n reaction chambers, and an idle region, the n reaction chambers are arranged in an array in the microwell array substrate, the idle region is arranged surrounding the n reaction chambers, the reaction chambers are configured to accommodate a sample to be measured, the shape of the orthogonal projection of the first main surface of the reaction chamber onto a first reference plane on which the first main surface of the reaction chamber is located is positive, the area of ​​the idle region is divided into n' virtual units, the shape of the orthogonal projection of the first main surface of the reaction chamber onto the first reference plane on which the first main surface of the reaction chamber is located and the shape of the orthogonal projection of the virtual unit onto the first reference plane are both regular N-gons, and the total volume V of the n reaction chambers satisfies Equation 1.

[0070]

number

[0071] In the microwell array chip according to the embodiments of the present disclosure, the total volume V of the n reaction chambers satisfies the above formula 1, and therefore the microwell array chip can efficiently utilize the area of ​​the microwell array substrate, thereby increasing the volume of a single reaction chamber, provided that the number of reaction chambers is sufficient, thereby improving the sensitivity of the microwell array chip and reducing the detection limit of the microwell array chip.

[0072] Microwell array chips, methods of using the same, and detection devices according to embodiments of the present disclosure will be described in detail below with reference to the drawings.

[0073] An embodiment of the present disclosure provides a microwell array chip. Figure 1 is a schematic plan view of a microwell array chip according to an embodiment of the present disclosure. Figure 2 is a schematic cross-sectional view of a microwell array chip according to an embodiment of the present disclosure.

[0074] 1 and 2, the microwell array chip 100 includes a microwell array substrate 110, which includes a first main surface 110A and a second main surface 110B arranged opposite to each other, n reaction chambers 120, and an idle region 130, the n reaction chambers 120 are arranged in an array within the microwell array substrate 110, the idle region 130 is arranged to surround the n reaction chambers 120, the reaction chambers 120 are configured to accommodate a sample to be measured, the shape of the orthogonal projection of the first main surface 110A of the reaction chambers 120 onto a first reference plane 201 on which the reaction chambers 120 are located is a regular N-gon, the area of ​​the idle region 130 is divided into n' virtual units 132, the shape of the orthogonal projection of the virtual units 132 onto the first reference plane 201 is the same as the shape of the orthogonal projection of the reaction chambers 120 onto the first reference plane 201, and the total volume V of the n reaction chambers 120 satisfies Equation 1.

[0075]

number

[0076] In the microwell array chip according to the embodiments of the present disclosure, the total volume V of the n reaction chambers satisfies the above formula 1, and therefore the microwell array chip can efficiently utilize the area of ​​the microwell array substrate, thereby increasing the volume of a single reaction chamber, provided that the number of reaction chambers is sufficient, thereby improving the sensitivity of the microwell array chip and reducing the detection limit of the microwell array chip.

[0077] In some examples, the total volume V of the n reaction chambers satisfies Equation 4.

[0078]

number

[0079] This enables the microwell array chip to efficiently utilize the area of ​​the microwell array substrate, improving the sensitivity of the microwell array chip and reducing the detection limit of the microwell array chip.

[0080] 1 and 2, the microwell array chip 110 further includes a support region 140 configured to provide a support structure. In this case, the total volume V of the n reaction chambers 120 satisfies Equation 2.

[0081]

number

[0082] In the microwell array chip according to this example, the support region may be provided in contact with a support structure (e.g., the support structure of the pedestal), so that there is a certain gap between the microwell array substrate and the pedestal, which allows liquid (e.g., the sample to be measured) to easily enter, thereby oil-sealing the microwell array substrate. Note that, although the support region adopts a different filling pattern in FIG. 1, the support region may be made of the same material as the other regions, i.e., the surface of the support region may be the same as the surface of the surrounding idle region. Of course, the embodiment of the present disclosure is not limited thereto, and the surface of the support region in contact with the support structure may be coated with other materials or structures or subjected to a surface treatment, so as to increase the friction force or other properties and to better contact the support structure.

[0083] 1 and 2, the microwell array chip 100 further includes a reaction region 150, which is disposed on the periphery of the support region 140, and the n reaction chambers 120 are disposed within the reaction region 150. As a result, no reaction chambers are disposed in the support region of the microwell array chip, and the reaction regions are disposed on the periphery of the support region, thereby making full use of the area of ​​the microwell array substrate.

[0084] For example, as shown in Figures 1 and 2, four support regions 140 are located on the four edges of the microwell array substrate 110 and are respectively located in the middle of the edges, and reaction regions 150 are provided in the middle region of the microwell array substrate 110 and are located around the support regions 140.

[0085] In addition, in the microwell array chip according to the embodiment of the present disclosure, the virtual units are provided to calculate the area of ​​the idle region and the area occupied by the reaction chamber as a unit, so that the virtual units are not only the same in shape as the reaction chambers, but also in the same arrangement form. For example, the interval between adjacent virtual units and reaction chambers, the interval between adjacent reaction chambers and reaction chambers, and the interval between adjacent virtual units may all be the same. For example, when reaction chambers are arranged according to a certain period to form a reaction chamber group (for example, to form a reaction chamber row or reaction chamber column), the virtual units may also be arranged according to a certain period to form a virtual group. For example, when reaction chambers are arranged regularly or staggered according to their shapes, the virtual units are also arranged regularly or staggered to fill the reaction region and the idle region in the entire microwell array substrate. However, when the microwell array chip includes a support region, the virtual units only need to avoid the support region, and the arrangement of the virtual units in the idle region is still the same as the arrangement of the reaction chambers.

[0086] In some examples, as shown in Figures 1 and 2, in the reaction area 150, the center distances of the orthogonal projections of two adjacent reaction chambers 120 onto the first reference plane 201 are equal. That is, the barriers between adjacent reaction chambers in the reaction area are equal and uniformly distributed. Thereby, the microwell array chip can ensure that the volume of liquid (e.g., the sample to be measured) entering each reaction chamber is the same, improving the accuracy of the reaction.

[0087] 1 and 2, the shape of the orthogonal projection of each reaction chamber 120 onto the first reference plane 201 is a regular hexagon, which allows the reaction chamber 120 to have a large volume and makes it easier for a liquid (e.g., a sample to be measured) to enter the reaction chamber. Of course, the embodiment of the present disclosure is not limited thereto, and the shape of the orthogonal projection of each reaction chamber 120 onto the first reference plane 201 may be a circle, a regular octagon, a regular pentagon, a square, a triangle, etc.

[0088] For example, the orthogonal projection of each reaction chamber onto the first reference plane may be a triangle, the small angles of each corner of which can break the surface tension of the test sample, thereby facilitating the entrance of the test sample into the reaction chamber.

[0089] In some examples, as shown in Figures 1 and 2, the shape of the orthogonal projection onto the first reference plane 201 on which the first main surface 110A of the reaction chamber 120 is located and the shape of the orthogonal projection onto the first reference plane 201 of the virtual unit 132 are both regular hexagons, and the total volume V of the n reaction chambers satisfies Equation 3.

[0090]

number

[0091] In the microwell array chip of this example, the shape of the orthogonal projection of each reaction chamber onto the first reference plane is a regular hexagon, and the total volume V of the n reaction chambers satisfies the above formula 3, so that the microwell array chip can efficiently utilize the area of ​​the microwell array substrate, thereby increasing the volume of a single reaction chamber, further improving the sensitivity of the microwell array chip, and reducing the detection limit of the microwell array chip.

[0092] In some examples, the size range of the spacing between adjacent reaction chambers 120 at the center line of adjacent reaction chambers 120 may be 20 to 40 microns, for example, 24 microns, 26 microns, 28 microns, 30 microns, 32 microns, 34 microns, or 36 microns.

[0093] In some examples, the depth of the reaction chamber 120 in a direction perpendicular to the first reference plane 201 may be 200 microns, 220 microns, 240 microns, 260 microns, 280 microns, or 300 microns.

[0094] In some examples, the number of reaction chambers 120 in one microwell array substrate 110 may be 8,000 to 100,000. This allows the microwell array chip to have higher detection accuracy.

[0095] In some examples, the number of reaction chambers 120 in one microwell array substrate 110 may be 8,000, 10,000, 20,000, 40,000, 60,000, 80,000, or 100,000.

[0096] FIG. 3A is a schematic plan view of a reaction chamber of another microwell array chip according to one embodiment of the present disclosure, and FIG. 3B is a schematic cross-sectional view of a reaction chamber of another microwell array chip according to one embodiment of the present disclosure.

[0097] As shown in Figures 3A and 3B, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A, and the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 is spaced apart from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201. In other words, the first hydrophobic layer 161 does not extend to the edge of the reaction chamber 120, which prevents the sample or reagent from remaining in an area outside the reaction chamber and allows the sample or reagent to easily enter the reaction chamber. Note that the first hydrophobic layer has hydrophobic and lipophilic properties, which allows the liquid (e.g., the sample to be measured) to easily enter each reaction chamber defined in the microwell array substrate.

[0098] For example, the material of the first hydrophobic layer may be a resin or silicon nitride, such as an epoxy resin. The first hydrophobic layer may be made of other suitable inorganic or organic materials, as long as it can ensure that the side of the first hydrophobic layer away from the microwell array substrate has hydrophobicity.

[0099] In some examples, as shown in Figures 3A and 3B, the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 on which the first major surface 110A is located includes a first opening 1610, the edge of which is spaced apart from the edge of the reaction chamber 120 on the first reference plane 201 and is located outside the edge of the reaction chamber 120 on the first reference plane 201.

[0100] In some examples, as shown in FIG. 3A, the shape of the orthogonal projection of the reaction chamber 120 onto the first reference plane 201 is a regular hexagon, the shape of the first opening 1610 is a regular hexagon, and the side length of the first opening 1610 is greater than the side length of the orthogonal projection of the reaction chamber 120 onto the first reference plane 201.

[0101] 3A and 3B, in the microwell array chip 100, the reaction chambers 120 penetrate the microwell array substrate 110 in a direction perpendicular to the microwell array substrate 110 (i.e., perpendicular to the first reference plane 201). That is, the reaction chambers 120 are through-holes that penetrate the microwell array substrate 110.

[0102] 3A and 3B, the microwell array chip 100 further includes a second hydrophobic layer 162 located on the second major surface 110B, the second hydrophobic layer 162 extending to the edge of the reaction chamber 120. Note that the second hydrophobic layer has hydrophobic and lipophilic properties, which can facilitate the ingress of liquid (e.g., a sample to be measured) into each reaction chamber defined in the microwell array substrate.

[0103] For example, the material of the second hydrophobic layer may be a resin or silicon nitride, such as an epoxy resin. The second hydrophobic layer may be made of other suitable inorganic or organic materials, as long as it can ensure that the side of the second hydrophobic layer away from the microwell array substrate has hydrophobicity.

[0104] 3A and 3B, the orthogonal projection of the second hydrophobic layer 162 onto the second reference plane 202 on which the second major surface 110B is located includes a second opening 1620, and an edge of the second opening 1620 overlaps with an edge of the reaction chamber 120 on the second reference plane 202. That is, the second hydrophobic layer is located just between adjacent reaction chambers.

[0105] In some examples, as shown in FIG. 3A, the shape of the orthogonal projection of the reaction chamber 120 onto the first reference plane 201 is a regular hexagon, the shape of the second opening 1620 is a regular hexagon, and the side length of the second opening 1620 is equal to the side length of the orthogonal projection of the reaction chamber 120 onto the first reference plane 201.

[0106] FIG. 4A is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure, and FIG. 4B is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure.

[0107] 4A and 4B, in the microwell array chip 100, the reaction chambers 120 penetrate the microwell array substrate 110 in a direction perpendicular to the microwell array substrate 110 (i.e., a direction perpendicular to the first reference plane 201). In other words, the reaction chambers 120 are through-holes that penetrate the microwell array substrate 110.

[0108] In some examples, as shown in Figures 4A and 4B, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A, and the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 is spaced apart from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201. In other words, the first hydrophobic layer 161 does not extend to the edge of the reaction chamber 120, which prevents the sample or reagent from remaining in an area outside the reaction chamber and allows the sample or reagent to easily enter the reaction chamber. Note that the first hydrophobic layer has hydrophobic and lipophilic properties, which allows the liquid (e.g., the sample to be measured) to easily enter each reaction chamber defined in the microwell array substrate.

[0109] For example, the material of the first hydrophobic layer may be a resin or silicon nitride, such as an epoxy resin. The first hydrophobic layer may be made of other suitable inorganic or organic materials, as long as it can ensure that the side of the first hydrophobic layer away from the microwell array substrate has hydrophobicity.

[0110] In some examples, as shown in Figures 4A and 4B, the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 on which the first major surface 110A is located includes a first opening 1610, the edge of which is spaced apart from the edge of the reaction chamber 120 on the first reference plane 201 and is located outside the edge of the reaction chamber 120 on the first reference plane 201.

[0111] 4A and 4B, the microwell array chip 100 further includes a second hydrophobic layer 162 located on the second main surface 110B, the second hydrophobic layer 162 spanning the reaction chamber 120, and the orthogonal projection of the second main surface 110B of the reaction chamber 120 onto the second reference plane 202 being within the orthogonal projection of the second hydrophobic layer 162 onto the second reference plane 202. That is, the microwell array chip can seal the side of the reaction chamber located on the second main surface with the second hydrophobic layer, thereby making the reaction chamber a blind hole.

[0112] In some examples, as shown in FIG. 4A, the shape of the orthogonal projection of the reaction chamber 120 onto the first reference plane 201 is a regular hexagon, and in this case, the shape of the first opening 1610 of the first hydrophobic layer 161 in the orthogonal projection onto the first reference plane 201 is also a regular hexagon, and the side length of the first opening 1610 is greater than the side length of the orthogonal projection of the reaction chamber 120 onto the first reference plane 201.

[0113] Figure 5 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. As shown in Figure 5, reaction chamber 120 is recessed from first main surface 110A into microwell array substrate 110, has body bottom 122 located within microwell array substrate 110, and the distance between body bottom 122 and first reference plane 201 is the thickness of microwell array substrate 110. That is, the reaction chamber is a blind hole.

[0114] 5, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A. The contact angle θ between the first hydrophobic layer 161 and the sample to be measured e is the critical angle θ of the reaction chamber 120 t0 is less than the critical angle θ e is the angle between the extension line of the side wall of the reaction chamber 120 and the tangent line of the surface of the sample to be measured that contacts the side wall of the reaction chamber 120. This allows the sample to be measured to enter the reaction chamber more easily in the microwell array chip.

[0115] In some examples, as shown in FIG. 5 , two adjacent reaction chambers 120 are provided in a first direction, a size in the first direction of a side wall between two adjacent reaction chambers 120 on a first reference plane 201 is Wtt, a size in the first direction of a side wall between two adjacent reaction chambers 120 on a second reference plane 202 is Wtb, a size in the first direction of an orthogonal projection of a body bottom 122 of the reaction chamber 120 onto the second reference plane 202 is St, and an included angle between the side wall of the reaction chamber 120 and a direction perpendicular to the second reference plane 202 is α t At this time, the critical angle θ of the reaction chamber t0 satisfies Equation 5.

[0116]

number

[0117] FIG. 6A is a schematic plan view of another microwell array chip according to one embodiment of the present disclosure, and FIG. 6B is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure.

[0118] 6A and 6B, reaction chamber 120 is recessed into microwell array substrate 110 from first main surface 110A and has a body bottom 122 located within microwell array substrate 110, and the distance between body bottom 122 and first reference plane 201 is the thickness of microwell array substrate 110. That is, the reaction chamber is a blind hole.

[0119] 6A and 6B, the body bottom 122 includes at least one exhaust hole 1220, and each exhaust hole 1220 penetrates the body bottom 122 in a direction perpendicular to the first reference plane 201. Thereby, when a liquid (e.g., a sample to be measured) enters the reaction chamber, the exhaust hole can be used to exhaust gas, thereby facilitating the liquid to enter the reaction chamber and increasing the speed at which the liquid enters the reaction chamber.

[0120] 6A and 6B, the main body bottom 122 includes one exhaust hole 1220, and the orthogonal projection of the second main surface 110B of the exhaust hole 1220 onto the second reference plane 202 is located at the center of the orthogonal projection of the main body bottom 122 onto the second reference plane 202. This allows air to be quickly exhausted from the exhaust hole of the reaction chamber, thereby increasing the speed at which liquid (e.g., a sample to be measured) enters the reaction chamber.

[0121] In some examples, as shown in Figures 6A and 6B, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A, and the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 is provided at a distance from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201. In other words, the first hydrophobic layer 161 does not extend to the edge of the reaction chamber 120, which prevents the sample or reagent from remaining in an area outside the reaction chamber and makes it easier for the sample or reagent to enter the reaction chamber. Note that the first hydrophobic layer has hydrophobic and lipophilic properties, which makes it easier for a liquid (e.g., a sample to be measured) to enter each reaction chamber defined in the microwell array substrate.

[0122] For example, the material of the first hydrophobic layer may be a resin or silicon nitride, such as an epoxy resin. The first hydrophobic layer may be made of other suitable inorganic or organic materials, as long as it can ensure that the side of the first hydrophobic layer away from the microwell array substrate has hydrophobicity.

[0123] In some examples, as shown in Figures 6A and 6B, the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 on which the first major surface 110A is located includes a first opening 1610, the edge of which is spaced apart from the edge of the reaction chamber 120 on the first reference plane 201 and is located outside the edge of the reaction chamber 120 on the first reference plane 201.

[0124] In some examples, as shown in Figures 6A and 6B, the microwell array chip 100 further includes a second hydrophobic layer 162 located on the second major surface 110B, the second hydrophobic layer 162 also including an exhaust opening 1625, which is in communication with the exhaust hole 1220.

[0125] FIG. 7A is a schematic plan view of another microwell array chip according to one embodiment of the present disclosure, and FIG. 7B is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure.

[0126] 7A and 7B, reaction chamber 120 is recessed from first main surface 110A into microwell array substrate 110 and has a body bottom 122 located within microwell array substrate 110, and the distance between body bottom 122 and first reference plane 201 is the thickness of microwell array substrate 110. That is, the reaction chamber is a blind hole.

[0127] 7A and 7B, the body bottom 122 includes a plurality of exhaust holes 1220, each of which penetrates the body bottom 122 in a direction perpendicular to the first reference plane 201, and the orthogonal projections of the plurality of exhaust holes 1220 onto the second reference plane 202 on which the second main surface 110B is located are arranged around the center of the orthogonal projection of the body bottom 122 onto the second reference plane 202. Thereby, when a liquid (e.g., a sample to be measured) enters the reaction chamber, the plurality of exhaust holes can be used simultaneously to exhaust gas, which makes it easier for the liquid to enter the reaction chamber and increases the speed at which the liquid enters the reaction chamber.

[0128] In some examples, as shown in FIG. 7A, the shape of the orthogonal projection of the reaction chamber 120 onto the second reference plane 202 is a regular hexagon, and in this case, the orthogonal projections of the six exhaust holes 1220 onto the second reference plane 202 are located at the six corners of the orthogonal projection of the reaction chamber 120 onto the second reference plane 202.

[0129] 7A and 7B, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A, and the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 is provided at a distance from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201. In other words, the first hydrophobic layer 161 does not extend to the edge of the reaction chamber 120, thereby preventing the sample or reagent from remaining in an area outside the reaction chamber and facilitating the sample or reagent to enter the reaction chamber.

[0130] In some examples, as shown in Figures 7A and 7B, the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 on which the first major surface 110A is located includes a first opening 1610, the edge of which is spaced apart from the edge of the reaction chamber 120 on the first reference plane 201 and is located outside the edge of the reaction chamber 120 on the first reference plane 201.

[0131] In some examples, as shown in Figures 7A and 7B, the microwell array chip 100 further includes a second hydrophobic layer 162 located on the second main surface 110B, the second hydrophobic layer 162 including a plurality of exhaust openings 1625, the plurality of exhaust openings 1625 being arranged in one-to-one correspondence with the plurality of exhaust openings 1220, and each exhaust opening 1625 being connected to a corresponding exhaust hole 1220.

[0132] FIG. 8A is a schematic plan view of another microwell array chip according to one embodiment of the present disclosure, and FIG. 8B is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure.

[0133] 8A and 8B, in microwell array chip 100, reaction chambers 120 penetrate microwell array substrate 110 in a direction perpendicular to microwell array substrate 110 (i.e., a direction perpendicular to first reference plane 201). In other words, reaction chambers 120 are through-holes that penetrate microwell array substrate 110.

[0134] 8A and 8B, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A, and the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 is provided at a distance from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201. In other words, the first hydrophobic layer 161 does not extend to the edge of the reaction chamber 120, thereby preventing the sample or reagent from remaining in an area outside the reaction chamber and facilitating the sample or reagent to enter the reaction chamber.

[0135] In some examples, as shown in Figures 8A and 8B, the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 on which the first major surface 110A is located includes a first opening 1610, the edge of which is spaced apart from the edge of the reaction chamber 120 on the first reference plane 201 and is located outside the edge of the reaction chamber 120 on the first reference plane 201.

[0136] 8A and 8B, the microwell array chip 100 further includes a dialysis membrane 170 and a second hydrophobic layer 162, the reaction chamber 120 penetrates the microwell array substrate 110 in a direction perpendicular to the first reference plane 201, the second hydrophobic layer 162 extends to the edge of the reaction chamber 120, and the dialysis membrane 170 spans the reaction chamber 120. The dialysis membrane thereby allows gas in the reaction chamber to be discharged but does not allow liquid in the reaction chamber to flow out, thereby increasing the speed at which the liquid (e.g., the sample to be measured) enters the reaction chamber and preventing the liquid from flowing out of the reaction chamber.

[0137] In some examples, as shown in Figures 8A and 8B, the orthogonal projection onto the second reference plane 202 on which the second main surface 110B of the reaction chamber 120 is located is within the orthogonal projection onto the second reference plane 202 of the dialysis membrane 170.

[0138] In some examples, as shown in Figures 8A and 8B, the dialysis membrane 170 is a flexible dialysis membrane. When the dialysis membrane is a flexible dialysis membrane, it can better contact with the liquid (e.g., the sample to be measured), thereby discharging the gas in the reaction chamber. Of course, the embodiment of the present disclosure is not limited thereto, and the dialysis membrane may be a thin film having a certain rigidity.

[0139] FIG. 9A is a schematic plan view of another microwell array chip according to one embodiment of the present disclosure, and FIG. 9B is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure.

[0140] 9A and 9B, in the microwell array chip 100, the reaction chambers 120 penetrate the microwell array substrate 110 in a direction perpendicular to the microwell array substrate 110 (i.e., a direction perpendicular to the first reference plane 201). In other words, the reaction chambers 120 are through-holes that penetrate the microwell array substrate 110.

[0141] 9A and 9B, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A, and the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 is provided at a distance from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201. In other words, the first hydrophobic layer 161 does not extend to the edge of the reaction chamber 120, thereby preventing the sample or reagent from remaining in an area outside the reaction chamber and facilitating the sample or reagent to enter the reaction chamber.

[0142] In some examples, as shown in Figures 9A and 9B, the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 on which the first major surface 110A is located includes a first opening 1610, the edge of which is spaced apart from the edge of the reaction chamber 120 on the first reference plane 201 and is located outside the edge of the reaction chamber 120 on the first reference plane 201.

[0143] 9A and 9B, the microwell array chip 100 further includes a dialysis membrane 170 and a second hydrophobic layer 162, where the reaction chambers 120 penetrate the microwell array substrate 110 in a direction perpendicular to the first reference plane 201, the second hydrophobic layer 162 extends to the edge of the reaction chambers 120, and the dialysis membrane 170 spans the reaction chambers 120. The dialysis membrane 170 is located on the side of the second hydrophobic layer 162 adjacent to the second main surface 110B.

[0144] FIG. 10A is a schematic plan view of another microwell array chip according to one embodiment of the present disclosure, and FIG. 10B is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure.

[0145] 10A and 10B, in microwell array chip 100, reaction chambers 120 penetrate microwell array substrate 110 in a direction perpendicular to microwell array substrate 110 (i.e., a direction perpendicular to first reference plane 201). In other words, reaction chambers 120 are through-holes that penetrate microwell array substrate 110.

[0146] 10A and 10B, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A, and the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 is provided at a distance from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201. In other words, the first hydrophobic layer 161 does not extend to the edge of the reaction chamber 120, thereby preventing the sample or reagent from remaining in an area outside the reaction chamber and facilitating the sample or reagent to enter the reaction chamber.

[0147] In some examples, as shown in Figures 10A and 10B, the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 on which the first main surface 110A is located includes a first opening 1610, the edge of which is spaced apart from the edge of the reaction chamber 120 on the first reference plane 201 and is located outside the edge of the reaction chamber 120 on the first reference plane 201.

[0148] 10A and 10B, the microwell array chip 100 further includes a dialysis membrane 170 and a second hydrophobic layer 162, where the reaction chambers 120 penetrate the microwell array substrate 110 in a direction perpendicular to the first reference plane 201, the second hydrophobic layer 162 extends to the edge of the reaction chambers 120, and the dialysis membrane 170 spans the reaction chambers 120. The dialysis membrane 170 is located on the side of the second hydrophobic layer 162 away from the second main surface 110B.

[0149] FIG. 11A is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure, FIG. 11B is a schematic oblique view of a reaction chamber of another microwell array chip according to one embodiment of the present disclosure, and FIG. 11C is a schematic oblique view of a reaction chamber of another microwell array chip according to one embodiment of the present disclosure.

[0150] 11A, 11B, and 11C, in microwell array chip 100, reaction chambers 120 penetrate microwell array substrate 110 in a direction perpendicular to microwell array substrate 110 (i.e., a direction perpendicular to first reference plane 201). In other words, reaction chambers 120 are through-holes that penetrate microwell array substrate 110.

[0151] 11A, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A, and the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 is provided at a distance from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201. In other words, the first hydrophobic layer 161 does not extend to the edge of the reaction chamber 120, thereby preventing the sample or reagent from remaining in an area outside the reaction chamber and facilitating the sample or reagent to enter the reaction chamber.

[0152] In some examples, as shown in FIG. 11A, the microwell array chip 100 further includes a second hydrophobic layer 162 located on the second major surface 110B, where the second hydrophobic layer 162 extends to the edge of the reaction chamber 120.

[0153] 11A, 11B, and 11C, the angle γ between the inner surface 126 of the reaction chamber 120 and the first main surface 110A is greater than 90 degrees. That is, the size of the portion of the reaction chamber 120 adjacent to the first main surface 110A is large. This makes it easier for liquid to enter the reaction chamber, thereby increasing the speed at which the liquid (e.g., the sample to be measured) enters the reaction chamber.

[0154] In some examples, as shown in FIG. 11B, the cross-sectional shape of the reaction chamber 120 cut at the first reference plane 201 on which the first main surface 110A is located is circular, the cross-sectional shape of the reaction chamber 120 cut at the second reference plane 202 on which the second main surface 110B is located is circular, and the diameter of the cross-section of the reaction chamber 120 cut at the first reference plane 201 is larger than the diameter of the cross-section of the reaction chamber 120 cut at the second reference plane 202.

[0155] In some examples, as shown in FIG. 11C, the cross-sectional shape of the reaction chamber 120 cut at the first reference plane 201 on which the first main surface 110A is located is a regular hexagon, the cross-sectional shape of the reaction chamber 120 cut at the second reference plane 202 on which the second main surface 110B is located is a regular hexagon, and the side length of the cross-section of the reaction chamber 120 cut at the first reference plane 201 is greater than the side length of the cross-section of the reaction chamber 120 cut at the second reference plane 202.

[0156] Figure 12A is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure, Figure 12B is a schematic oblique view of a reaction chamber of another microwell array chip according to one embodiment of the present disclosure, and Figure 12C is a schematic oblique view of a reaction chamber of another microwell array chip according to one embodiment of the present disclosure.

[0157] 12A, 12B, and 12C, in microwell array chip 100, reaction chambers 120 penetrate microwell array substrate 110 in a direction perpendicular to microwell array substrate 110 (i.e., a direction perpendicular to first reference plane 201). In other words, reaction chambers 120 are through-holes that penetrate microwell array substrate 110.

[0158] 12A, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A, and the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 is provided at a distance from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201. In other words, the first hydrophobic layer 161 does not extend to the edge of the reaction chamber 120, thereby preventing the sample or reagent from remaining in an area outside the reaction chamber and facilitating the sample or reagent to enter the reaction chamber.

[0159] In some examples, as shown in FIG. 12A, the microwell array chip 100 further includes a second hydrophobic layer 162 located on the second major surface 110B, where the second hydrophobic layer 162 extends to the edge of the reaction chamber 120.

[0160] 12A, 12B, and 12C, the inner surface 126 of the reaction chamber 120 includes a first sub-surface 1261 and a second sub-surface 1262 in a direction perpendicular to the first reference plane 201, the second sub-surface 1262 is located on the side of the first sub-surface 1261 away from the first main surface 110A, the angle between the first sub-surface 1261 and the first main surface 110A is greater than 90 degrees, and the angle between the second sub-surface 1262 and the second main surface 110B is greater than 90 degrees. In other words, the size of the part of the reaction chamber 120 adjacent to the first main surface 110A is large, the size of the part of the reaction chamber 120 adjacent to the second main surface 110B is large, and the size of the middle part of the reaction chamber 120 is small. This allows the reaction chamber to be easily filled with liquid on the one hand and easily discharge gas on the other hand, thereby further increasing the rate at which liquid (eg, a sample to be measured) enters the reaction chamber.

[0161] In some examples, as shown in FIG. 12B , the cross-sectional shape of the reaction chamber 120 cut at the first reference plane 201 on which the first main surface 110A is located is circular, the cross-sectional shape of the reaction chamber 120 cut at the second reference plane 202 on which the second main surface 110B is located is circular, the cross-sectional shape of the reaction chamber 120 cut at the third reference plane 203 parallel to the first reference plane 201 and the second reference plane 202 and located at the first reference plane 201 and the second reference plane 202 is circular, and both the diameter of the cross-section of the reaction chamber 120 cut at the first reference plane 201 and the diameter of the cross-section of the reaction chamber 120 cut at the second reference plane 202 are larger than the diameter of the cross-section of the reaction chamber 120 cut at the third reference plane 203 parallel to the first reference plane 201 and the second reference plane 202 and located at the first reference plane 201 and the second reference plane 202.

[0162] In some examples, as shown in FIG. 12C, the cross-sectional shape of the reaction chamber 120 cut at the first reference plane 201 on which the first main surface 110A is located is a regular hexagon, the cross-sectional shape of the reaction chamber 120 cut at the second reference plane 202 on which the second main surface 110B is located is a regular hexagon, the cross-sectional shape of the reaction chamber 120 cut at the third reference plane 203 parallel to the first reference plane 201 and the second reference plane 202 and located on the first reference plane 201 and the second reference plane 202 is a regular hexagon, and both the side lengths of the cross-section of the reaction chamber 120 cut at the first reference plane 201 and the second reference plane 202 are greater than the side length of the cross-section of the reaction chamber 120 cut at the third reference plane 203.

[0163] Figure 13A is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure, Figure 13B is a schematic oblique view of a reaction chamber of another microwell array chip according to one embodiment of the present disclosure, and Figure 13C is a schematic oblique view of a reaction chamber of another microwell array chip according to one embodiment of the present disclosure.

[0164] 13A, 13B, and 13C, in microwell array chip 100, reaction chambers 120 penetrate microwell array substrate 110 in a direction perpendicular to microwell array substrate 110 (i.e., a direction perpendicular to first reference plane 201). In other words, reaction chambers 120 are through-holes that penetrate microwell array substrate 110.

[0165] 13A, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A, and the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 is provided at a distance from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201. In other words, the first hydrophobic layer 161 does not extend to the edge of the reaction chamber 120, thereby preventing the sample or reagent from remaining in an area outside the reaction chamber and facilitating the sample or reagent to enter the reaction chamber.

[0166] In some examples, as shown in FIG. 13A, the microwell array chip 100 further includes a second hydrophobic layer 162 located on the second major surface 110B, where the second hydrophobic layer 162 extends to the edge of the reaction chamber 120.

[0167] 13A, 13B, and 13C, the inner surface 126 of the reaction chamber 120 includes a first sub-surface 1261, a second sub-surface 1262, and a third sub-surface 1263 in a direction perpendicular to the first reference plane 201, the second sub-surface 1262 being located on the side of the first sub-surface 1261 that is away from the first major surface 110A, and the third sub-surface 1263 being located on the side of the second sub-surface 1262 that is away from the first sub-surface 1261. The angle between the first sub-surface 1261 and the first major surface 110A is greater than 90 degrees, the plane of the second sub-surface 1262 is perpendicular to the first reference plane 201, and the angle between the third sub-surface 1263 and the second major surface 110B is greater than 90 degrees. In this case, the size of the part of the reaction chamber 120 adjacent to the first main surface 110A is large, the size of the part of the reaction chamber 120 adjacent to the second main surface 110B is large, and the size of the middle part of the reaction chamber 120 is small. This makes it easy for the reaction chamber to receive liquid on the one hand and to discharge gas on the other hand, thereby further increasing the speed at which the liquid (e.g., the sample to be measured) enters the reaction chamber. Also, since the plane of the second sub-surface is perpendicular to the first reference plane, the middle part of the reaction chamber facilitates the storage of liquid.

[0168] In some examples, as shown in FIG. 13B, the cross-sectional shape of the reaction chamber 120 cut at the first reference plane 201 on which the first main surface 110A is located is circular, the cross-sectional shape of the reaction chamber 120 cut at the second reference plane 202 on which the second main surface 110B is located is circular, the cross-sectional shape of the reaction chamber 120 cut at the third reference plane 203 parallel to the first reference plane 201 and the second reference plane 202 and located at the first reference plane 201 and the second reference plane 202 is circular, and both the diameter of the cross-section of the reaction chamber 120 cut at the first reference plane 201 and the diameter of the cross-section of the reaction chamber 120 cut at the second reference plane 202 are larger than the diameter of the cross-section of the reaction chamber 120 cut at the third reference plane 203 parallel to the first reference plane 201 and the second reference plane 202 and located at the first reference plane 201 and the second reference plane 202.

[0169] In some examples, as shown in FIG. 13C, the cross-sectional shape of the reaction chamber 120 cut at the first reference plane 201 on which the first main surface 110A is located is a regular hexagon, the cross-sectional shape of the reaction chamber 120 cut at the second reference plane 202 on which the second main surface 110B is located is a regular hexagon, the cross-sectional shape of the reaction chamber 120 cut at the third reference plane 203 parallel to the first reference plane 201 and the second reference plane 202 and located on the first reference plane 201 and the second reference plane 202 is a regular hexagon, and both the side lengths of the cross-section of the reaction chamber 120 cut at the first reference plane 201 and the side lengths of the cross-section of the reaction chamber 120 cut at the second reference plane 202 are greater than the side lengths of the cross-section of the reaction chamber 120 cut at the third reference plane 203.

[0170] Figure 14A is a schematic cross-sectional view of another microwell array chip according to one embodiment of the present disclosure, Figure 14B is a schematic oblique view of a reaction chamber of another microwell array chip according to one embodiment of the present disclosure, and Figure 14C is a schematic oblique view of a reaction chamber of another microwell array chip according to one embodiment of the present disclosure.

[0171] 14A, 14B, and 14C, in microwell array chip 100, reaction chambers 120 penetrate microwell array substrate 110 in a direction perpendicular to microwell array substrate 110 (i.e., a direction perpendicular to first reference plane 201). In other words, reaction chambers 120 are through-holes that penetrate microwell array substrate 110.

[0172] 14A, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A, and the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 is provided at a distance from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201. In other words, the first hydrophobic layer 161 does not extend to the edge of the reaction chamber 120, thereby preventing the sample or reagent from remaining in an area outside the reaction chamber and facilitating the sample or reagent to enter the reaction chamber.

[0173] In some examples, as shown in FIG. 14A, the microwell array chip 100 further includes a second hydrophobic layer 162 located on the second major surface 110B, where the second hydrophobic layer 162 extends to the edge of the reaction chamber 120.

[0174] 14A, 14B, and 14C, the inner surface 126 of the reaction chamber 120 includes a first sub-surface 1261, a second sub-surface 1262, and a third sub-surface 1263 in a direction perpendicular to the first reference plane 201, the second sub-surface 1262 being located on the side of the first sub-surface 1261 that is away from the first major surface 110A, and the third sub-surface 1263 being located on the side of the second sub-surface 1262 that is away from the first sub-surface 1261. The angle between the first sub-surface 1261 and the first major surface 110A is greater than 90 degrees, the second sub-surface 1262 is a circular arc surface that is concave toward the microwell array substrate 110, and the angle between the third sub-surface 1263 and the second major surface 110B is greater than 90 degrees. In this case, the size of the portion of the reaction chamber 120 adjacent to the first main surface 110A is large, the size of the portion of the reaction chamber 120 adjacent to the second main surface 110B is large, and the size of the middle portion of the reaction chamber 120 is small. This allows the reaction chamber to easily allow liquid to enter on the one hand and to easily discharge gas on the other hand, thereby further increasing the speed at which liquid (e.g., a sample to be measured) enters the reaction chamber. In addition, since the second sub-surface is an arc surface that is concave toward the microwell array substrate, the middle portion of the reaction chamber facilitates the storage of liquid.

[0175] In some examples, as shown in FIG. 14B , the cross-sectional shape of the reaction chamber 120 cut at the first reference plane 201 on which the first main surface 110A is located is circular, the cross-sectional shape of the reaction chamber 120 cut at the second reference plane 202 on which the second main surface 110B is located is circular, the cross-sectional shape of the reaction chamber 120 cut at the third reference plane 203 parallel to the first reference plane 201 and the second reference plane 202 and located at the first reference plane 201 and the second reference plane 202 is circular, and both the diameter of the cross-section of the reaction chamber 120 cut at the first reference plane 201 and the diameter of the cross-section of the reaction chamber 120 cut at the second reference plane 202 are larger than the diameter of the cross-section of the reaction chamber 120 cut at the third reference plane 203 parallel to the first reference plane 201 and the second reference plane 202 and located at the first reference plane 201 and the second reference plane 202.

[0176] In some examples, as shown in FIG. 14C, the cross-sectional shape of the reaction chamber 120 cut at the first reference plane 201 on which the first main surface 110A is located is a regular hexagon, the cross-sectional shape of the reaction chamber 120 cut at the second reference plane 202 on which the second main surface 110B is located is a regular hexagon, the cross-sectional shape of the reaction chamber 120 cut at the third reference plane 203 parallel to the first reference plane 201 and the second reference plane 202 and located on the first reference plane 201 and the second reference plane 202 is a regular hexagon, and both the side lengths of the cross-section of the reaction chamber 120 cut at the first reference plane 201 and the side lengths of the cross-section of the reaction chamber 120 cut at the second reference plane 202 are greater than the side lengths of the cross-section of the reaction chamber 120 cut at the third reference plane 203.

[0177] Figure 15 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. As shown in Figure 15, in microwell array chip 100, reaction chambers 120 penetrate microwell array substrate 110 in a direction perpendicular to microwell array substrate 110 (i.e., a direction perpendicular to first reference plane 201). In other words, reaction chambers 120 are through-holes that penetrate microwell array substrate 110. A first hydrophilic film 181 and a second hydrophilic film 182 are provided on the inner surface 126 of the reaction chamber 120, the first hydrophilic film 181 and the second hydrophilic film 182 are provided adjacent to each other in a direction perpendicular to the first reference plane 201, and the surface of the first hydrophilic film 181 away from the inner surface 126 of the reaction chamber 120 is an arcuate surface convex toward the central axis of the reaction chamber 120, and the surface of the second hydrophilic film 182 away from the inner surface 126 of the reaction chamber 120 is an arcuate surface convex toward the central axis of the reaction chamber 120. As a result, on the one hand, the first hydrophilic film and the second hydrophilic film can not only facilitate the entry of liquid (e.g., the sample to be measured) into the reaction chamber, but also facilitate the storage of liquid; on the other hand, the surface of the first hydrophilic film away from the inner surface of the reaction chamber is an arc-shaped surface that is convex toward the central axis of the reaction chamber, so the size of the portion adjacent to the first main surface of the reaction chamber is large, making it easier for liquid to enter the reaction chamber; similarly, the surface of the second hydrophilic film away from the inner surface of the reaction chamber is arc-shaped, so the size of the portion adjacent to the second main surface of the reaction chamber is large, which is advantageous for discharging gas; and also, a recess (i.e., the adjacent portion of the first hydrophilic film and the second hydrophilic film) that is concave toward the microwell array substrate can be further formed between the first hydrophilic film and the second hydrophilic film, thereby making it easier to store liquid.

[0178] In some examples, as shown in FIG. 15, the inner surface 126 of the reaction chamber 120 is flat, and the thickness of the first hydrophilic film 181 in a direction perpendicular to the inner surface 126 varies, such that the surface of the first hydrophilic film 181 that faces away from the inner surface 126 of the reaction chamber 120 is an arcuate surface, and the thickness of the second hydrophilic film 182 in a direction perpendicular to the inner surface 126 varies, such that the surface of the second hydrophilic film 182 that faces away from the inner surface of the reaction chamber 120 is an arcuate surface.

[0179] Figure 16 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. As shown in Figure 16, in microwell array chip 100, reaction chambers 120 penetrate microwell array substrate 110 in a direction perpendicular to microwell array substrate 110 (i.e., a direction perpendicular to first reference plane 201). In other words, reaction chambers 120 are through-holes that penetrate microwell array substrate 110. A first hydrophilic film 181 and a second hydrophilic film 182 are provided on the inner surface 126 of the reaction chamber 120, the first hydrophilic film 181 and the second hydrophilic film 182 are arranged adjacent to each other in a direction perpendicular to the first reference plane 201, the inner surface 126 of the reaction chamber 120 includes a first sub-surface 1261 and a second sub-surface 1262 in the direction perpendicular to the first reference plane 201, the second sub-surface 1262 is located on the side of the first sub-surface 1261 away from the first main surface 110A, and the first sub-surface 1261 is convex toward the central axis of the reaction chamber 120, such that the surface of the first hydrophilic film 181 away from the inner surface 126 of the reaction chamber 120 is an arcuate surface, and the second sub-surface 1262 is convex toward the central axis of the reaction chamber 120, such that the surface of the second hydrophilic film 182 away from the inner surface 126 of the reaction chamber 120 is an arcuate surface. As a result, on the one hand, the first hydrophilic film and the second hydrophilic film can not only facilitate the entry of liquid (e.g., the sample to be measured) into the reaction chamber, but also facilitate the storage of liquid; on the other hand, the surface of the first hydrophilic film away from the inner surface of the reaction chamber is an arc-shaped surface that is convex toward the central axis of the reaction chamber, so the size of the portion adjacent to the first main surface of the reaction chamber is large, making it easier for liquid to enter the reaction chamber; similarly, the surface of the second hydrophilic film away from the inner surface of the reaction chamber is arc-shaped, so the size of the portion adjacent to the second main surface of the reaction chamber is large, which is advantageous for discharging gas; and also, a recess (i.e., the adjacent portion of the first hydrophilic film and the second hydrophilic film) that is concave toward the microwell array substrate can be further formed between the first hydrophilic film and the second hydrophilic film, thereby making it easier to store liquid.

[0180] Fig. 17 is a structural schematic diagram of a microwell array chip according to an embodiment of the present disclosure. As shown in Fig. 17, the microwell array chip 100 further includes a first packaging film 191 and a second packaging film 192, the first packaging film 191 being located on the side of the first main surface 110A that is away from the second main surface 110B, the second packaging film 192 being located on the side of the second main surface 110B that is away from the first main surface 110A, and the first packaging film 191 and the second packaging film 192 being attached to the microwell array substrate 110 by electrostatic or colloidal adhesion. This makes the microwell array chip easy to use, and after injecting a liquid (e.g., a sample to be measured) into the microwell array substrate, the first packaging film and the second packaging film can be attached to the microwell array substrate by electrostatic adsorption or colloidal adhesion.

[0181] 17, the microwell array chip 100 further includes a first hydrophobic layer 161 and a second hydrophobic layer 162, the first hydrophobic layer 161 being provided on the first main surface 110A, the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 being spaced apart from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201, and the second hydrophobic layer 162 being located on the second main surface 110B. At this time, the first packaging film 191 is located on the side of the first hydrophobic layer 161 that is away from the second main surface 110B, and the second packaging film 192 is located on the side of the second hydrophobic layer 162 that is away from the first main surface 110A. Thereby, the first packaging film and the second packaging film can well package the microwell array substrate. In addition, the first hydrophobic layer has hydrophobic and lipophilic properties, which can facilitate the entry of liquid (eg, a sample to be measured) into each reaction chamber defined in the microwell array substrate.

[0182] For example, the material of the first hydrophobic layer and the second hydrophobic layer may be a resin or silicon nitride, such as an epoxy resin. The first hydrophobic layer and the second hydrophobic layer may also be made of other suitable inorganic or organic materials.

[0183] 18 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. As shown in FIG. 18, the microwell array chip 100 further includes a photocurable oil 210 located at an opening position adjacent to the first main surface 110A of the reaction chamber 120. This allows the microwell array chip to package the reaction chamber with the photocurable oil. Alternatively, the photocurable oil may be first injected into the opening position adjacent to the first main surface of the reaction chamber, and then the photocurable oil may be cured by a photocuring process, thereby packaging the reaction chamber.

[0184] 18, the microwell array chip 100 further includes a first hydrophobic layer 161 provided on the first main surface 110A, and the orthogonal projection of the first hydrophobic layer 161 onto the first reference plane 201 is provided at a distance from the orthogonal projection of the reaction chamber 120 onto the first reference plane 201. In other words, the first hydrophobic layer 161 does not extend to the edge of the reaction chamber 120, thereby preventing the sample or reagent from remaining in an area outside the reaction chamber and facilitating the sample or reagent to enter the reaction chamber.

[0185] In some examples, as shown in FIG. 18, the photocurable oil 210 includes a boss structure 212 that is disposed in contact with the first major surface 110A and is located on the side of the first hydrophobic layer 161 that is closer to the central axis of the reaction chamber 120.

[0186] Fig. 19 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. As shown in Fig. 19, microwell array substrate 110 is a flexible substrate. In this case, microwell array substrate 110 may include a first sub-flexible microwell array substrate 115A and a second sub-flexible microwell array substrate 115B, where first sub-flexible microwell array substrate 115A and second sub-flexible microwell array substrate 115B are arranged in contact with each other, and a plurality of reaction chambers 120 are formed between first sub-flexible microwell array substrate 115A and second flexible microwell array substrate 115B.

[0187] FIG. 20 is a schematic plan view of another microwell array chip according to an embodiment of the present disclosure. As shown in FIG. 20, the microwell array substrate 110 includes a liquid introduction channel 128, n reaction chambers 120 communicate with the liquid introduction channel 128, and a unidirectional membrane 129 is provided between each reaction chamber 120 and the liquid introduction channel 128. As a result, the microwell array chip can inject a liquid (e.g., a sample to be measured) into the reaction chamber through the liquid introduction channel, and the unidirectional membrane can prevent the test sample in the reaction chamber from returning to the liquid introduction channel, thereby preventing crosstalk between different reaction chambers. Of course, the embodiment of the present disclosure is not limited to this, and a unidirectional membrane does not have to be provided between each reaction chamber and the liquid introduction channel.

[0188] 20, the liquid introduction channel 128 includes a main liquid introduction channel 1282 and n liquid introduction branch channels 1284 each communicating with the main liquid introduction channel 1282, and the n liquid introduction branch channels 1284 are provided in one-to-one correspondence with the n reaction chambers 120. Thereby, the microwell array chip can inject liquid (e.g., a sample to be measured) into the reaction chambers via the main liquid introduction channel and the branch liquid introduction channels.

[0189] In some examples, as shown in FIG. 20 , n liquid inlet branch channels 1284 are provided on both sides of the main liquid inlet channel 1282, and the multiple liquid inlet branch channels 1284 located on the first side of the main liquid inlet channel 1282 and the multiple liquid inlet branch channels 1284 located on the second side of the main liquid inlet channel 1282 are arranged in a staggered pattern.

[0190] Fig. 21 is a schematic plan view of another microwell array chip according to an embodiment of the present disclosure. As shown in Fig. 21, the microwell array substrate 110 includes a liquid introduction channel 128, which includes a plurality of main liquid introduction channels 1282 arranged in parallel, and n liquid introduction branch channels 1284, and on both sides of each main liquid introduction channel 1282, a plurality of liquid introduction branch channels 1284 communicating with the main liquid introduction channel 1282 are provided, and the n liquid introduction branch channels 1284 are provided in one-to-one correspondence with the n reaction chambers 120. As a result, the microwell array chip can inject liquid (e.g., a sample to be measured) into the reaction chambers via the liquid introduction channel.

[0191] 22 is a schematic plan view of another microwell array chip according to an embodiment of the present disclosure. As shown in FIG. 22, the microwell array substrate 110 includes a liquid introduction channel 128, which includes a plurality of sub-liquid introduction channels 1285 that communicate with each other, and each of the sub-liquid introduction channels 1285 communicates with a plurality of reaction chambers 120. The height of the sub-liquid introduction channel 1285 is greater than the height of the plurality of reaction chambers 120 connected thereto. This allows liquid to flow from the sub-liquid introduction channel into the reaction chamber by gravity. The above "height" refers to the distance between the bottom of the sub-liquid introduction channel or the reaction chamber and the second main surface of the microwell array substrate.

[0192] In some examples, the heights of the multiple sub-liquid introduction channels 1285 are different. Since the heights of the multiple sub-liquid introduction channels are different, the microwell array chip can use gravity to cause liquid (e.g., a sample to be measured) to flow from a higher sub-liquid introduction channel to a lower sub-liquid introduction channel, thereby injecting the liquid into all of the reaction chambers. Note that the above phrase "the heights of the multiple sub-liquid introduction channels are different" means that the distances between the bottoms of the sub-liquid introduction channels and the second main surface of the microwell array substrate are different.

[0193] 22, the multiple sub liquid introduction channels 1285 are arranged in sequence, and the heights of the multiple sub liquid introduction channels 1285 decrease in sequence along the arrangement direction of the multiple sub liquid introduction channels 1285. Of course, the embodiments of the present disclosure are not limited thereto, and the heights of the multiple liquid introduction channels 1285 may decrease in sequence from the middle toward both sides.

[0194] Figure 23 is a schematic plan view of another microwell array chip according to an embodiment of the present disclosure. As shown in Figure 23, the microwell array substrate 110 includes a liquid introduction channel 128, and the shape of the orthogonal projection of liquid introduction channel 128 onto first reference plane 201 on which first main surface 110A of microwell array substrate 110 is located is a broken curve. This allows the microwell array chip to directly inject liquid (e.g., a sample to be measured) into the reaction chambers via the liquid introduction channel.

[0195] 24 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. As shown in FIG. 24, the microwell array chip 100 further includes a first substrate 310 and a second substrate 320, the first substrate 220 being located on one side of the microwell array substrate 110 and spaced apart from the first main surface 110A of the microwell array substrate 110, the second substrate 320 being located on the side of the microwell array substrate 110 that is away from the first substrate 310, the second substrate 320 including a heating electrode 325, the orthogonal projection of the heating electrode 325 onto the first reference plane 201 on which the first main surface 110A of the microwell array substrate 110 is located overlapping with the orthogonal projection of at least a part of the n reaction chambers 120 onto the first reference plane 201. As a result, the microwell array chip can heat at least a part of the n reaction chambers by the heating electrode, and thus the heating function can be integrated into the microwell array chip.

[0196] 24, the orthogonal projection of the heating electrode 325 onto the first reference plane 201 on which the first main surface 110A of the microwell array substrate 110 is located overlaps with the orthogonal projection of the n reaction chambers 120 onto the first reference plane 201. This allows the microwell array chip to simultaneously heat the n reaction chambers by the heating electrode.

[0197] 24, the first substrate 310 includes a first base substrate 311 and a third hydrophobic layer 312, the third hydrophobic layer 312 being located on a side of the first base substrate 311 adjacent to the second substrate 320. The third hydrophobic layer 312 thereby has hydrophobic and lipophilic properties, which can facilitate the entry of a liquid (e.g., a sample to be measured) into each reaction chamber 120 defined in the microwell array substrate 110.

[0198] For example, the material of the third hydrophobic layer may be resin or silicon nitride, such as epoxy resin. The third hydrophobic layer may be made of other suitable inorganic or organic materials, as long as it can ensure that the side of the third hydrophobic layer facing the second substrate has hydrophobicity.

[0199] 24, the first substrate 310 and the second substrate 310 can form a receiving space 340 by a sealant 330, the microwell array substrate 110 is disposed in the receiving space 340, and the first substrate 310 further includes at least one sample inlet 315, which penetrates the first base substrate 311 and the third hydrophobic layer 312 and communicates with the receiving space 340. Thus, the microwell array chip can inject liquid into the microwell array substrate through the sample inlet.

[0200] 24, the first substrate 310 further includes at least one sample outlet 317, which penetrates the first base substrate 311 and the third hydrophobic layer 312 and communicates with the receiving space 340. Thereby, the microwell array chip can discharge liquid through the sample outlet.

[0201] 24, the second substrate 320 further includes a second base substrate 321, a control electrode 322, a first insulating layer 323, and a second insulating layer 324, the control electrode 322 being located on the second base substrate 321, the first insulating layer 323 being located on the side of the control electrode 322 that is remote from the second base substrate 321, the first insulating layer 323 including a connection hole 323H, the connection hole 323H exposing at least a part of the control electrode 322, the heating electrode 325 being located on the side of the first insulating layer 323 that is remote from the second base substrate 321 and connected to the control electrode 322 via the connection hole 323H, the second insulating layer 324 being located on the side of the heating electrode 325 that is remote from the first insulating layer 323, and the microwell array substrate 100 being located on the second insulating layer 324. Thereby, the microwell array chip can drive the heating electrode to generate heat by applying a voltage to the heating electrode by the control electrode. Meanwhile, since the second insulating layer is located on the side of the heating electrode away from the first insulating layer, the second insulating layer can protect the heating electrode and prevent the corrosion of water and oxygen, thereby extending the service life of the heating electrode. The second insulating layer can also play the role of insulation and planarization.

[0202] For example, the first insulating layer 323 and the second insulating layer 324 may be made of the same material or different materials. The first insulating layer 323 and the second insulating layer 324 may be made of an inorganic insulating material such as silicon oxide, silicon nitride, or silicon oxynitride, or may be made of an organic insulating material such as resin or polyimide.

[0203] 24, the microwell array chip 100 further includes a photosensitive sensor 380 located on the side of the second substrate 320 away from the first substrate 310 and configured to detect light emitted by the reaction chambers 120 of the microwell array substrate 110. The microwell array chip can thereby determine whether a reaction occurs in the reaction chambers and the extent of the reaction through the photosensitive sensor, and the microwell array chip also integrates the photosensitive sensor within the chip, thereby further improving the degree of integration.

[0204] FIG. 25 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. As shown in FIG. 25, the microwell array chip 100 further includes a first substrate 310 and a second substrate 320, and the first substrate 220 is located on one side of the microwell array substrate 110 and is spaced apart from the first main surface 110A of the microwell array substrate 110. The microwell array chip 100 includes a plurality of microwell array substrates 110 arranged in a direction parallel to the first substrate 310 or the second substrate 320. As a result, the microwell array chip can achieve high-throughput detection by providing a plurality of microwell array chips. Note that in the embodiment of the present disclosure, the planar layout structure of the plurality of microwell array chips is not particularly limited. For example, the plurality of microwell array substrates may be arranged in a matrix, and the plurality of microwell array substrates may be provided surrounding one microwell array substrate.

[0205] In some examples, as shown in Figure 25, when the microwell array chip includes multiple microwell array substrates 110, the second substrate 320 may include a heating electrode 325, and the orthogonal projection of the heating electrode 325 onto the first reference plane 201 on which the first main surface 110A of the microwell array substrate 110 is located overlaps with the orthogonal projection onto the first reference plane 201 of at least some of the n reaction chambers 120 in at least one microwell array substrate 110. This allows the microwell array chip to heat at least some of the n reaction chambers in at least one microwell array substrate by the heating electrode, thereby integrating a heating function into the microwell array chip. Note that the microwell array chip may also be provided with multiple heating electrodes, and the multiple heating electrodes are provided in one-to-one correspondence with the multiple microwell array substrates.

[0206] 25, the first substrate 310 includes a first base substrate 311 and a third hydrophobic layer 312, the third hydrophobic layer 312 being located on a side of the first base substrate 311 adjacent to the second substrate 320. The third hydrophobic layer 312 thereby has hydrophobic and lipophilic properties, which can facilitate the entry of a liquid (e.g., a sample to be measured) into each reaction chamber 120 defined in the microwell array substrate 110.

[0207] For example, the material of the third hydrophobic layer may be resin or silicon nitride, such as epoxy resin. The third hydrophobic layer may be made of other suitable inorganic or organic materials, as long as it can ensure that the side of the third hydrophobic layer facing the second substrate has hydrophobicity.

[0208] 25, the first substrate 310 and the second substrate 310 can form a receiving space 340 by a sealant 330, the microwell array substrate 110 is disposed within the receiving space 340, and the first substrate 310 further includes at least one sample inlet 315, which penetrates the first base substrate 311 and the third hydrophobic layer 312 and communicates with the receiving space 340. Thus, the microwell array chip can inject liquid into the multiple microwell array substrates through the sample inlet.

[0209] 25, the first substrate 310 further includes at least one sample outlet 317, which penetrates the first base substrate 311 and the third hydrophobic layer 312 and communicates with the receiving space 340. Thereby, the microwell array chip can discharge liquid through the sample outlet.

[0210] 25, the second substrate 320 further includes a second base substrate 321, a control electrode 322, a first insulating layer 323, and a second insulating layer 324, the control electrode 322 being located on the second base substrate 321, the first insulating layer 323 being located on the side of the control electrode 322 that is remote from the second base substrate 321, the first insulating layer 323 including a connection hole 323H, the connection hole 323H exposing at least a part of the control electrode 322, the heating electrode 325 being located on the side of the first insulating layer 323 that is remote from the second base substrate 321 and connected to the control electrode 322 via the connection hole 323H, the second insulating layer 324 being located on the side of the heating electrode 325 that is remote from the first insulating layer 323, and the microwell array substrate 100 being located on the second insulating layer 324. Thereby, the microwell array chip can drive the heating electrode to generate heat by applying a voltage to the heating electrode by the control electrode. Meanwhile, since the second insulating layer is located on the side of the heating electrode away from the first insulating layer, the second insulating layer can protect the heating electrode and prevent the corrosion of water and oxygen, thereby extending the service life of the heating electrode. The second insulating layer can also play the role of insulation and planarization.

[0211] For example, the first insulating layer 323 and the second insulating layer 324 may be made of the same material or different materials. The first insulating layer 323 and the second insulating layer 324 may be made of an inorganic insulating material such as silicon oxide, silicon nitride, or silicon oxynitride, or may be made of an organic insulating material such as resin or polyimide.

[0212] 25, the microwell array chip 100 further includes a photosensitive sensor 380 located on the side of the second substrate 320 away from the first substrate 310 and configured to detect light emitted by the reaction chambers 120 of the microwell array substrate 110. The microwell array chip can thereby determine whether a reaction occurs in the reaction chambers and the extent of the reaction through the photosensitive sensor, and the microwell array chip also integrates the photosensitive sensor within the chip, thereby further improving the degree of integration.

[0213] 25 achieves high-throughput detection by arranging multiple microwell array substrates in a direction parallel to the first substrate or the second substrate. However, embodiments of the present disclosure are not limited thereto, and the microwell array chip may achieve high-throughput detection by arranging multiple microwell array substrates in a direction perpendicular to the first substrate or the second substrate.

[0214] 26 is a schematic cross-sectional view of another microwell array chip according to an embodiment of the present disclosure. As shown in FIG. 26, the microwell array chip 100 includes a microwell array substrate 110, which includes a first main surface 110A and a second main surface 110B that are provided opposite to each other, n reaction chambers 120, and an idle region 130. The n reaction chambers 120 are provided in an array in the microwell array substrate 110, and each reaction chamber 120 is configured to accommodate a sample to be measured. The microwell array chip 100 further includes an electrically induced hydrophilicity change layer 190 provided on an inner surface 126 of the reaction chamber 120, which is connected to an external circuit by a conductive structure (e.g., a conductor), and can change the hydrophilicity (e.g., decrease hydrophilicity or change from hydrophilic to hydrophobic) by passing an electric current through the layer, thereby automatically discharging the liquid in the reaction chamber 120.

[0215] In some examples, the microwell array substrate in the microwell array chip may be manufactured from an avidin material, thereby enabling reuse.

[0216] In some examples, the microwell array substrate in the microwell array chip may be fabricated from a material that bypasses the fluorescence excitation band, thereby preventing interference with fluorescence detection and improving detection accuracy.

[0217] Furthermore, the total volume V of the n reaction chambers 120 in the microwell array chips shown in Figures 1 to 26 above can all satisfy Equation 1, thereby allowing the area of ​​the microwell array substrate to be utilized efficiently, thereby increasing the volume of a single reaction chamber, and further improving the sensitivity of the microwell array chip while reducing the detection limit of the microwell array chip.

[0218]

number

[0219] At least one embodiment of the present disclosure further provides a detection device. FIG. 27 is a schematic diagram of a detection device according to one embodiment of the present disclosure. As shown in FIG. 27, the detection device 500 includes a microwell array chip 100 according to any one of the above examples. The microwell array chip can efficiently utilize the area of ​​the microwell array substrate, thereby increasing the volume of a single reaction chamber, and further improving the sensitivity of the microwell array chip while reducing the detection limit of the microwell array chip. Thus, the detection device including the microwell array chip has the same high sensitivity and low detection limit.

[0220] Fig. 28 is a schematic diagram of another detection device according to an embodiment of the present disclosure. As shown in Fig. 28, the detection device 500 further includes a first casing 510 and a second casing 520, where the first casing 510 is located on one side of the microwell array chip 100 and is spaced apart from the first main surface 110A, and the second casing 520 is located on the side of the microwell array chip 100 that is away from the first casing 510 and is spaced apart from the second main surface 110B. The distance between the second main surface 110B and the second casing 520 is equal to or greater than the thickness of the microwell array chip 100. This allows the detection device to facilitate the entry of liquid into the microwell array chip.

[0221] Fig. 29 is a schematic plan view of a second casing of a detection device according to an embodiment of the present disclosure. As shown in Fig. 28, second casing 520 includes a support structure 525, which includes a first platform portion 525A and a second platform portion 525B, the height of second platform portion 525B being greater than the height of first platform portion 525A, first platform portion 525A being configured to contact the bottom surface of microwell array chip 100, and second platform portion 525B being configured to contact the side surface of microwell array chip 100. Thereby, the detection device can support the microwell array chip by the first platform portion and position the microwell array chip by the second platform portion, thereby improving detection accuracy.

[0222] In some examples, as shown in FIG. 29, the shape of the orthogonal projection of the first main surface 110A of the first platform portion 525A onto the first reference plane 201 on which it is located includes an arc triangle, and the shape of the orthogonal projection of the first main surface 110A of the second platform portion 525B onto the first reference plane 201 (for the first reference plane, see FIG. 27) on which it is located includes a semicircle, the base side connected to the semicircle of the arc triangle is a straight line, and the remaining two sides of the arc triangle are arcs.

[0223] In some examples, as shown in FIG. 29, the second casing 520 further includes a positioning ring 528 configured to be placed in contact with the side of the microwell array chip 100, thereby further positioning the microwell array chip and improving detection accuracy.

[0224] At least one embodiment of the present disclosure further provides a method of using a microwell array chip, the microwell array chip including a microwell array substrate, the method including the following steps S101 to S103.

[0225] Step S101: A measurement sample is injected into a microwell array substrate.

[0226] Step S102: The sample to be measured is packaged in a microwell array substrate, the microwell array substrate includes a first main surface and a second main surface arranged opposite to each other, the microwell array substrate includes n reaction chambers and an idle area, the n reaction chambers are arranged in an array in the microwell array substrate and configured to accommodate the sample to be measured, the shape of the orthogonal projection onto a first reference plane on which the first main surface of the reaction chamber is located is a regular N-gon, the idle area is arranged to surround the n reaction chambers, the area of ​​the idle area is divided into n' virtual units, the shape of the orthogonal projection onto the first reference plane of the virtual units is the same as the shape of the orthogonal projection of the reaction chambers onto the first reference plane, and the total volume V of the n reaction chambers satisfies Equation 1.

[0227]

number

[0228] In the method of using a microwell array chip according to an embodiment of the present disclosure, the total volume V of the n reaction chambers satisfies the above formula 1, and therefore the method of use can efficiently utilize the area of ​​the microwell array substrate, thereby increasing the volume of a single reaction chamber, provided that the number of reaction chambers is sufficient, and further improving the sensitivity of the microwell array chip and reducing the detection limit of the microwell array chip.

[0229] In some examples, the total volume V of the n reaction chambers satisfies Equation 4.

[0230]

number

[0231] This enables the microwell array chip to efficiently utilize the area of ​​the microwell array substrate, improving the sensitivity of the microwell array chip and reducing the detection limit of the microwell array chip.

[0232] In some examples, the microwell array chip further comprises a support region configured to provide a support structure, wherein the total volume V of the n reaction chambers satisfies Equation 2.

[0233]

number

[0234] In the method of using the microwell array chip according to this example, the support region may be provided in contact with a support structure (e.g., the support structure of the pedestal), thereby leaving a certain gap between the microwell array substrate and the pedestal, making it easier for the sample to be measured to enter, and thereby enabling the microwell array substrate to be oil-sealed.

[0235] In some examples, the shape of the orthogonal projection onto a first reference plane on which the first main surface of the reaction chamber is located and the shape of the orthogonal projection onto the first reference plane of the virtual unit are both regular hexagons, and the total volume V of the n reaction chambers satisfies Equation 3.

[0236]

number

[0237] In the microwell array chip of this example, the shape of the orthogonal projection of each reaction chamber onto the first reference plane is a regular hexagon, and the total volume V of the n reaction chambers satisfies the above formula 3, so that the microwell array chip can efficiently utilize the area of ​​the microwell array substrate, thereby increasing the volume of a single reaction chamber, and further improving the sensitivity of the microwell array chip while reducing the detection limit of the microwell array chip.

[0238] In some examples, the spacing between adjacent reaction chambers at the center line of the adjacent reaction chambers may range from 20 to 40 microns, e.g., 24 microns, 26 microns, 28 microns, 30 microns, 32 microns, 34 microns, or 36 microns.

[0239] In some examples, the depth of the reaction chamber in a direction perpendicular to the first reference plane may be 200 microns, 220 microns, 240 microns, 260 microns, 280 microns, or 300 microns.

[0240] In some examples, the number of reaction chambers in one microwell array substrate may be 8000 to 100000. This allows the microwell array chip to have higher detection accuracy.

[0241] In some examples, the number of reaction chambers in a single microwell array substrate may be 8,000, 10,000, 20,000, 40,000, 60,000, 80,000, or 100,000.

[0242] In some examples, packaging the sample to be measured in the microwell array substrate includes, after injecting the sample to be measured into the microwell array substrate, attaching a first packaging film to the side of the first main surface away from the second main surface by electrostatic or colloidal, and attaching a second packaging film to the side of the second main surface away from the first main surface, so that the microwell array chip can easily package the microwell array substrate, with high efficiency.

[0243] In some examples, the step of packaging the sample to be measured in the microwell array substrate includes the steps of: coating a photocurable oil at an opening position adjacent to the first main surface of the reaction chamber after injecting the sample to be measured into the microwell array substrate; and curing the photocurable oil using ultraviolet light. Thus, the microwell array chip can package the microwell array substrate using the photocurable oil and the photocuring process, which has the advantages of being simple, efficient, and having good packaging effect.

[0244] In some examples, when the microwell array substrate uses the flexible substrate shown in Figure 19, the microwell array substrate may include a first sub-flexible microwell array substrate and a second sub-flexible microwell array substrate, and a plurality of sample flow paths are included between the first sub-flexible microwell array substrate and the second sub-flexible microwell array substrate. Injecting the sample to be measured into the microwell array substrate includes injecting the sample to be measured into the plurality of sample flow paths.

[0245] For example, the sample to be measured may be injected into the multiple sample flow paths by vacuum suction, pumping, or the like.

[0246] In some examples, packaging the sample to be measured in the microwell array substrate includes using a roller to partition each sample flow path to form a plurality of reaction chambers and seal the plurality of reaction chambers. For example, the roller has a heating function and can heat-bond the first sub-flexible microwell array substrate and the second flexible microwell array substrate at regular intervals, thereby partitioning each sample flow path to form a plurality of reaction chambers.

[0247] There are a few points that need to be explained.

[0248] (1) In the drawings of the disclosed embodiments, only the structures related to the embodiments of the present disclosure are shown, and other structures may be referred to standard designs.

[0249] (2) Unless inconsistent, features of the same and different embodiments of the present disclosure may be combined with each other.

[0250] Although the specific embodiments of the present disclosure have been described above, the scope of protection of the present disclosure is not limited thereto, and all modifications and replacements that a person skilled in the art can easily conceive without departing from the technical scope disclosed in the present disclosure should be included in the scope of protection of the present disclosure. Therefore, the scope of protection of the present disclosure should be in accordance with the scope of protection of the claims. [Explanation of symbols]

[0251] 100 Microwell Array Chips 110 Microwell array substrate 110A 1st main surface 110B 2nd main surface 115A First sub-flexible microwell array substrate 115B Second sub-flexible microwell array substrate 120 Reaction Chamber 122 Bottom of main body 126 Inner surface 128 Liquid introduction channel 129 Unidirectional membrane 130 Idle Area 132 Virtual Units 140 Support area 150 reaction regions 161 First hydrophobic layer 162 Second hydrophobic layer 170 Dialysis Membrane 181 1st hydrophilic membrane 182 Second hydrophilic membrane 190 Electrically Induced Hydrophilic Change Layer 191 First package membrane 192 Second package membrane 201 1st reference plane 202 Second reference plane 203 Third reference plane 210 Photohardened oil 212 Boss Structure 220 First Board 310 First board 311 First base substrate 312 Third hydrophobic layer 315 Sample inlet 317 Sample outlet 320 Second Board 321 Second base substrate 322 Control Electrode 323 First insulating layer 323H Connection hole 324 Second insulating layer 325 Heating electrode 330 Sealant 340 Storage Space 380 Photosensitive Sensor 500 Detection Device 510 First casing 520 Second Casing 525 Support structure 525A First Platform Section 525B Second Platform Section 528 yen range 1220 Exhaust Vent 1261 1st sub surface 1262 Second sub surface 1263 3rd sub-surface 1282 Liquid introduction main channel 1284 Liquid introduction branch channel 1285 Sub-liquid introduction channel 1610 First opening 1620 2nd opening 1625 Exhaust opening

Claims

1. A microwell array chip, comprising: a microwell array substrate including a first main surface and a second main surface disposed opposite to each other; n reaction chambers arranged in an array within the microwell array substrate, configured to accommodate a measurement target sample, the shape of which, when orthogonally projected onto a first reference plane on which the first main surface is located, is a regular N-sided polygon; an idle region surrounding the n reaction chambers; The area of ​​the idle region is divided into n′ virtual units, the shape of the orthogonal projection of the virtual units onto the first reference plane is the same as the shape of the orthogonal projection of the reaction chambers onto the first reference plane, and the total volume V of the n reaction chambers satisfies Equation 1; [0010] where (1-α) is the confidence level and S ｃｈｉｐ is the area of ​​the microwell array substrate, h is the depth of the reaction chamber in a direction perpendicular to the first reference plane, N is a positive integer of 3 or more, and X is 1 / 2 the size of the spacing between adjacent reaction chambers on the central connecting line of the adjacent reaction chambers.

2. a support region configured to provide a support structure; The total volume V of the n reaction chambers satisfies Equation 2: [0025] In the formula, S ｓｕｐｐｏｒｔ The microwell array chip of claim 1 , wherein is the area of ​​the support region.

3. The method further includes a reaction area provided around the support area, The microwell array chip of claim 2 , wherein the n reaction chambers are located in the reaction region.

4. The microwell array chip of claim 3 , wherein in the reaction region, the center distances of the orthogonal projections of two adjacent reaction chambers onto the first reference plane are equal.

5. The shape of the orthogonal projection of the first main surface of the reaction chamber onto a first reference plane is a regular hexagon, and the total volume V of the n reaction chambers satisfies Equation 3: [0030] The microwell array chip of any one of claims 1 to 4, wherein the value of X ranges from 10 to 20 microns and the value of h ranges from 190 to 320 microns.

6. The microwell array chip of any one of claims 1 to 4, wherein the value of n is in the range of 8,000 to 100,000.

7. The total volume V of the n reaction chambers is calculated by the following formula 4: [0045] The microwell array chip according to any one of claims 1 to 4, which satisfies the above.

8. a first hydrophobic layer located on the first major surface; The microwell array chip according to any one of claims 1 to 4, wherein an orthogonal projection of the first hydrophobic layer onto the first reference plane is spaced apart from an orthogonal projection of the reaction chamber onto the first reference plane.

9. further comprising a second hydrophobic layer located on the second major surface; the second hydrophobic layer extends to an edge of the reaction chamber; or The microwell array chip of claim 8, wherein the reaction chamber penetrates the microwell array substrate in a direction perpendicular to the first reference plane, the second hydrophobic layer spans the reaction chamber, and the orthogonal projection onto the second reference plane on which the second main surface of the reaction chamber is located is within the orthogonal projection of the second hydrophobic layer onto the second reference plane.

10. the reaction chamber is recessed from the first main surface into the microwell array substrate and has a body bottom located within the microwell array substrate, the distance between the body bottom and the first reference plane being less than a thickness of the microwell array substrate; 9. The microwell array chip of claim 8, wherein a contact angle between the first hydrophobic layer and the sample to be measured is less than a critical angle of the reaction chamber, the critical angle being an angle between an extension of a side wall of the reaction chamber and a tangent to a surface of the sample to be measured that contacts the side wall of the reaction chamber.

11. The microwell array chip of claim 10 , wherein the bottom of the body includes at least one exhaust hole, each of the exhaust holes penetrating the bottom of the body in a direction perpendicular to the first reference plane.

12. Dialysis membrane and a second hydrophobic layer, The microwell array chip of claim 8, wherein the reaction chamber penetrates the microwell array substrate in a direction perpendicular to the first reference plane, the second hydrophobic layer extends to an edge of the reaction chamber, and the dialysis membrane spans the reaction chamber.

13. the angle between the inner surface of the reaction chamber and the first main surface is greater than 90 degrees; an inner surface of the reaction chamber includes a first sub-surface and a second sub-surface in a direction perpendicular to the first reference plane, the second sub-surface being located on a side of the first sub-surface away from the first major surface, the included angle between the first sub-surface and the first major surface being greater than 90 degrees, and the included angle between the second sub-surface and the second major surface being greater than 90 degrees; or the inner surface of the reaction chamber includes a first sub-surface, a second sub-surface, and a third sub-surface in a direction perpendicular to the first reference plane, the second sub-surface being located on a side of the first sub-surface away from the first main surface, the third sub-surface being located on a side of the second sub-surface away from the first sub-surface, an included angle between the first sub-surface and the first main surface being greater than 90 degrees, a plane on which the second sub-surface is located is perpendicular to the first reference plane, and an included angle between the third sub-surface and the second main surface being greater than 90 degrees; or 5. The microwell array chip of any one of claims 1 to 4, wherein the inner surface of the reaction chamber includes a first sub-surface, a second sub-surface, and a third sub-surface in a direction perpendicular to the first reference plane, the second sub-surface is located on the side of the first sub-surface away from the first main surface, the third sub-surface is located on the side of the second sub-surface away from the first sub-surface, the included angle between the first sub-surface and the first main surface is greater than 90 degrees, the second sub-surface is an arcuate surface that is concave toward the microwell array substrate, and the included angle between the third sub-surface and the second main surface is greater than 90 degrees.

14. 5. The microwell array chip of any one of claims 1 to 4, wherein a first hydrophilic film and a second hydrophilic film are provided on an inner surface of the reaction chamber, the first hydrophilic film and the second hydrophilic film are provided adjacent to each other in a direction perpendicular to the first reference plane, the surface of the first hydrophilic film away from the inner surface of the reaction chamber is an arcuate surface that is convex toward a central axis of the reaction chamber, and the surface of the second hydrophilic film away from the inner surface of the reaction chamber is an arcuate surface that is convex toward the central axis of the reaction chamber.

15. 5. The microwell array chip according to any one of claims 1 to 4, wherein the shape of the orthogonal projection of the reaction chamber onto the first reference plane is virtually one of a circle, a regular hexagon, and a regular octagon.

16. The microwell array chip according to any one of claims 1 to 4, wherein the shape of the orthogonal projection of said reaction chamber onto said first reference plane is virtually a triangle.

17. a first packaging film located on a side of the first major surface away from the second major surface; a second packaging membrane located on a side of the second major surface away from the first major surface, The microwell array chip according to any one of claims 1 to 4, wherein the first packaging film and the second packaging film are attached onto the microwell array substrate by electrostatic or colloidal adhesion.

18. a photocurable oil in an open position adjacent the first major surface of the reaction chamber; 9. The microwell array chip of claim 8, wherein the photocurable oil includes a boss structure, the boss structure being provided in contact with the first main surface and located on a side of the first hydrophobic layer adjacent to a central axis of the reaction chamber.

19. The microwell array chip according to any one of claims 1 to 4, wherein the microwell array substrate is a flexible substrate.

20. The microwell array substrate comprises: Further comprising a liquid introduction channel; 5. The microwell array chip according to claim 1, wherein the n reaction chambers communicate with the liquid introduction channel, and a unidirectional membrane is provided between each of the reaction chambers and the liquid introduction channel.

21. a first substrate located on one side of the microwell array substrate and spaced apart from the first main surface; a second substrate located on a side of the microwell array substrate away from the first substrate, The microwell array chip of any one of claims 1 to 4, wherein the second substrate includes a heating electrode, and an orthogonal projection of the heating electrode onto the first reference plane overlaps with an orthogonal projection of at least a portion of the n reaction chambers onto the first reference plane.

22. A detection device comprising the microwell array chip according to any one of claims 1 to 4.

23. a first casing located on one side of the microwell array chip and spaced apart from the microwell array chip; a second casing located on a side of the microwell array chip away from the first casing and spaced apart from the microwell array chip; the distance between the microwell array chip and the second casing is equal to or greater than the thickness of the microwell array chip; the second casing includes a support structure, the support structure including a first platform portion and a second platform portion, the height of the second platform portion being greater than the height of the first platform portion, the first platform portion configured to contact a bottom surface of the microwell array chip, and the second platform portion configured to contact a side surface of the microwell array chip; a shape of an orthogonal projection of the first platform portion onto the first main surface includes an arc triangle, a shape of an orthogonal projection of the second platform portion onto the first main surface includes a semicircle, a base of the arc triangle connected to the semicircle is a straight line, and the remaining two sides of the arc triangle are arcs, 23. The detection apparatus of claim 22, wherein the second casing further comprises a positioning ring configured to be placed in contact with a side of the microwell array chip.

24. A method of using a microwell array chip, the microwell array chip comprising a microwell array substrate, the method of using the microwell array chip comprising: Injecting a sample to be measured into the microwell array substrate; packaging the sample to be measured in the microwell array substrate; The microwell array substrate includes n reaction chambers and an idle region, the n reaction chambers are arranged in an array in the microwell array substrate and are configured to accommodate a sample to be measured, the shape of the orthogonal projection of the first main surface of the reaction chamber onto a first reference plane on which the first main surface is located is a regular N-gon, the virtual idle region is arranged to surround the n reaction chambers, the area of ​​the virtual idle region is divided into n′ virtual units, the shape of the orthogonal projection of the virtual unit onto the first reference plane is the same as the shape of the orthogonal projection of the reaction chamber onto the first reference plane, and the total volume V of the n reaction chambers satisfies Equation 1, [0050] where (1-α) is the confidence level and S ｃｈｉｐ is the area of ​​the microwell array substrate, h is the depth of the reaction chamber in a direction perpendicular to the first reference plane, N is a positive integer of 3 or more, and X is 1 / 2 the size of the spacing between adjacent reaction chambers on a central connecting line of the adjacent reaction chambers.

25. The step of packaging the measurement sample in the microwell array substrate includes:

25. A method of using the microwell array chip of claim 24, comprising the steps of: after injecting a sample to be measured into the microwell array substrate, attaching a first packaging membrane to the side of the first main surface facing away from the second main surface by static electricity or colloids; and attaching a second packaging membrane to the side of the second main surface facing away from the first main surface.

Images