Microfluidic chip and testing method thereof

Abstract

The present invention relates to the technical field of microfluidics, and in particular to a microfluidic chip and a testing method thereof. The microfluidic chip comprises: a body, a first sample loading hole formed in the body, and sample separating channels communicated with the first sample loading hole, wherein: an anti-blocking region is provided between the sample separating channels and the first sample loading hole, and comprises a first surface formed by being raised in a direction moving away from the lower surface of the body, and a second surface connected to the lower surface; sample separating inlets of the sample separating channels are formed in the second surface; at least two first protrusions formed by protruding toward the lower surface are provided on the first surface, wherein first transition channels are formed between adjacent first protrusions; second transition channels are formed between the at least two first protrusions and the second surface, separately, and the second transition channels are adjacently connected to the sample separating inlets. By means of the arrangement of double-stage transition channels, the present invention can ensure that a blood sample can be smoothly separated under capillary action while filtering the blood sample.

Description

A microfluidic chip and its detection method

[0001] Priority application

[0002] This application claims priority to Chinese Invention Patent Application No. [2024208715919], filed on April 24, 2024, entitled "[A Microfluidic Chip with Anti-blocking Function]", which is incorporated herein by reference in its entirety. Technical Field

[0003] This invention relates to the field of microfluidics, specifically to a microfluidic chip and its detection method. Background Technology

[0004] Patent application CN112221545A discloses a multi-channel microfluidic sample loading device and its application. This sample loading device (also referred to as a chip) includes: a body; a sample loading port located on the surface of the body for receiving liquid samples; multiple capillary sample loading channels spaced apart inside the body, with the first end of each capillary sample loading channel communicating with the sample loading port; and multiple puncture tubes spaced apart on one side of the body and extending into the body, with the first end of each puncture tube inside the body communicating with the second end of each capillary sample loading channel.

[0005] The aforementioned sample loading device can automatically separate samples through the loading orifice and capillary sampling channel under high-speed centrifugation. However, in practical applications, the sample may become blocked at the entrance of the capillary sampling channel, resulting in incomplete or uneven sample separation, which affects the accuracy of the test results.

[0006] Therefore, there is an urgent need for a microfluidic chip with anti-clogging function. Summary of the Invention

[0007] The purpose of this invention is to provide a microfluidic chip with anti-clogging function, which partially solves or alleviates the above-mentioned deficiencies in the prior art and can effectively alleviate the clogging problem of microfluidic chips during the sample separation process.

[0008] In order to solve the technical problems mentioned above, the present invention adopts the following technical solution: The purpose of the present invention is to provide a microfluidic chip that partially solves or alleviates the above-mentioned deficiencies in the prior art and can alleviate the clogging problem in the sample separation process.

[0009] To solve the aforementioned technical problems, the present invention specifically adopts the following technical solution: A first aspect of the present invention is to provide a microfluidic chip, comprising:

[0010] ontology;

[0011] The first sample dispensing port has an opening located on the upper surface of the body.

[0012] A first number of sampling channels, each of which is connected to the first sample dispensing port, and an anti-clogging area is provided between the sampling channels and the first sample dispensing port; wherein, the anti-clogging area includes:

[0013] A transition zone is provided along at least a portion of the edge of the first sample dispensing hole and located on the lower surface of the body. The transition zone is formed by raising the height by a first height in a direction away from the lower surface. The transition zone includes a first surface formed by the elevation and a second surface connected to the lower surface. The sample dispensing inlet of the sample dispensing channel is provided on the second surface, and the first surface is a quasi-annular region surrounding at least a portion of the edge of the first sample dispensing hole.

[0014] At least two first protrusions are arranged at intervals on the first surface, protruding toward the lower surface. A second number of first transition channels are sequentially formed between adjacent first protrusions among the at least two first protrusions, and the width of the first transition channel gradually increases along the direction from its first end to its second end.

[0015] The at least two first protrusions respectively form a third number of second transition channels with the second surface, and at least one of the second transition channels is adjacent to and connected to the sample dispensing inlet; wherein, the sample located in the first sample feeding hole can enter the sample dispensing channel by sequentially passing through the first transition channel and the second transition channel under the action of driving force;

[0016] The first protrusion is configured to satisfy the following rule: the first quantity is less than or equal to the second quantity, and the second quantity is less than the third quantity.

[0017] In some embodiments, the third quantity is greater than or equal to twice the second quantity.

[0018] In some embodiments, at least one pair of the second transition channels are respectively disposed on both sides of one of the sample inlets.

[0019] In some embodiments, the height of the first protrusion is 0.05-0.20 mm.

[0020] In some embodiments, a second protrusion is further provided between at least two adjacent first protrusions, and the third and fourth sides of the second protrusion cooperate with the first protrusions on both sides to form a first supplementary channel and a second supplementary channel.

[0021] In some embodiments, the first inlet of the first transition channel is greater than or equal to the width of the second transition channel, and the width of the second transition channel is the same as or similar to the width of the sampling channel.

[0022] In some embodiments, it further includes a patch layer for sealing at least one channel.

[0023] In some embodiments, a flow-limiting region is provided at the second end of the sampling channel. The flow-limiting region includes a flow-limiting channel whose width gradually increases from its first end to its second end. The second end of the flow-limiting channel is connected to a communicating channel disposed on the body through a first through hole. Correspondingly, an auxiliary region is provided on the bonding layer. The auxiliary region includes a first number of first auxiliary regions, and the first auxiliary region is a first groove adapted to the shape of the sampling channel. The first groove is connected to the sampling channel and forms a complete sampling channel. A first absorbent layer is provided on the first groove. The first absorbent layer is made of a hydrophilic material so that the complete sampling channel has a first hydrophilic ability. The bonding layer is connected to the flow-limiting region and forms a complete flow-limiting region so that the complete flow-limiting region has a second hydrophilic ability, and the first hydrophilic ability is greater than or equal to the second hydrophilic ability.

[0024] For example, in some embodiments, the region in the bonding layer corresponding to the flow-limiting zone can be configured as a hydrophobic layer to enhance the flow-limiting effect of the flow-limiting zone.

[0025] In some embodiments, the bonding layer is a hydrophobic film.

[0026] In some embodiments, it further includes: a second sample feeding hole, wherein the opening of the second sample feeding hole is disposed on the upper surface;

[0027] A sample dispensing channel, wherein a first end of the sample dispensing channel is connected to a second sample dispensing hole, and a second end of the sample dispensing channel is connected to a communicating channel through a second through hole; wherein the first through hole is located above the second through hole;

[0028] The width of the sample addition channel is greater than the width of the sample separation channel, and the width of the connecting channel is less than the width of the connecting channel.

[0029] The present invention also provides a microfluidic detection method, comprising the following steps:

[0030] S100, providing a microfluidic chip as described in any of the above embodiments; wherein the complete sample dispensing channel is configured to have a first hydrophilicity, the sample dispensing channel is configured to have a third hydrophilicity, and the third hydrophilicity is greater than or equal to the first hydrophilicity; the width of the sample dispensing channel is greater than the width of the sample dispensing channel, and the length of the sample dispensing channel is less than the length of the sample dispensing channel;

[0031] S101, a first sample is added to the first sample feeding hole and a second sample is added to the second sample feeding hole respectively; wherein, the first sample can enter the sample dispensing channel sequentially through the first transition channel and the second transition channel under capillary action, and stop at the flow restriction zone, and the first sample needs to overcome the first resistance f1 applied by the flow restriction zone to continue entering the connecting channel; the second sample can enter the sample feeding channel under capillary action, and needs to overcome the second resistance f2 to continue entering the connecting channel, and the second resistance f2 is less than the first resistance f1;

[0032] S102, the microfluidic chip is centrifuged using a first centrifugal force f3, and the first centrifugal force f3 satisfies a first centrifugation rule: f1-f3>λ1, f3-f2>λ2, where λ1 is a first set value and λ2 is a second set value;

[0033] S103, the microfluidic chip is further centrifuged using a second centrifugal force f4, and the second centrifugal force f4 satisfies the second centrifugation rule: f4-f1>λ3, where λ3 is a first set value. Attached Figure Description

[0034] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. The elements or parts in the drawings are not necessarily drawn to scale. Obviously, the drawings described below are some embodiments of the present invention, and those skilled in the art can obtain other drawings based on these drawings without any creative effort.

[0035] Figure 1 is a schematic diagram of the anti-blocking area structure in the first exemplary embodiment of this application;

[0036] Figure 2 is a three-dimensional structural diagram of the anti-blocking area in the first exemplary embodiment of this application;

[0037] Figure 3 is a schematic diagram of the lower surface structure of the anti-blocking chip in the first exemplary embodiment of this application;

[0038] Figure 4 is a three-dimensional structural diagram of the anti-blocking chip in the first exemplary embodiment of this application;

[0039] Figure 5 is a schematic diagram of the anti-blocking chip in the first exemplary embodiment of this application;

[0040] Figure 6 is a first perspective view of the chip in the second example embodiment of this application;

[0041] Figure 7 is a schematic diagram of the chip structure in the second example embodiment of this application;

[0042] Figure 8 is a schematic diagram of the back structure of the chip in the second example embodiment of this application;

[0043] Figure 9 is a second perspective view of the chip in the second example embodiment of this application;

[0044] Figure 10 is a cross-sectional view of the chip in the second example embodiment of this application;

[0045] Figure 11 is a partial cross-sectional structural diagram of the bonding layer in the third exemplary embodiment of this application;

[0046] Figure 12 is a cross-sectional structural diagram of the detection card in an exemplary embodiment of this application.

[0047] Summary of reference numerals in the attached diagram: 10 is the main body, 11 is the first sample feeding port, 12 is the sample dispensing channel, 13 is the anti-clogging area, 131 is the transition zone, 131a is the first surface, 131b is the second surface, 132 is the first protrusion, 133 is the second protrusion, I is the first transition channel, I1 is the first inlet, I2 is the second inlet, II is the second transition channel, 14 is the waste liquid pool, L1 is the first side, L2 is the second side, L3 is the third side, L4 is the fourth side, L5 is the fifth side, 15 is the flow restriction zone, 15a is the flow restriction channel, 15b is the first through hole, 16 is the connecting channel, 17 is the second sample feeding port, 18 is the sample feeding channel, 19 is the second through hole, 20 is the bonding layer, 21 is the auxiliary area, 211 is the first auxiliary area, 212 is the second auxiliary area, 30 is the detection card, and 31 is the reaction chamber. Detailed Implementation

[0048] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0049] In this document, suffixes such as "module," "part," or "unit" used to denote elements are used only for the purpose of illustrative purposes and have no specific meaning in themselves. Therefore, "module," "part," or "unit" may be used interchangeably.

[0050] In this document, the terms "upper," "lower," "inner," "outer," "front," "rear," "one end," and "the other end," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing the present invention and for simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the present invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0051] In this document, unless otherwise explicitly specified and limited, the terms "installed," "equipped with," "connected," etc., should be interpreted broadly. For example, "connection" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection, a direct connection, or an indirect connection through an intermediate medium; it can be a connection within two components. Those skilled in the art can understand the specific meaning of the above terms in this invention based on the specific circumstances.

[0052] In this document, "and / or" includes any and all combinations of one or more of the listed related items.

[0053] In this article, "multiple" means two or more, that is, it includes two, three, four, five, etc.

[0054] As used in this specification, the term "about" typically means + / -5% of the value, more typically + / -4%, more typically + / -3%, more typically + / -2%, even more typically + / -1%, and even more typically + / -0.5%.

[0055] In this specification, certain embodiments may be disclosed in a range-bound format. It should be understood that this "range-bound" description is merely for convenience and brevity and should not be construed as a rigid limitation on the disclosed range. Therefore, the description of a range should be considered as having specifically disclosed all possible subranges and the individual numerical values ​​within those ranges. For example, a description of the range 1-6 should be considered as having specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and the individual numbers within those ranges, such as 1, 2, 3, 4, 5, and 6. This rule applies regardless of the breadth of the range.

[0056] In this article, the width of the channel refers to the length in a direction perpendicular or approximately perpendicular to the flow direction of the sample (or liquid sample, liquid) within the chip, while the height is also referred to as the thickness of the chip.

[0057] In this article, "hydrophilic substances" refers to molecules with polar groups that have a strong affinity for water, attract water molecules, and are easily wetted by water. "Hydrophilicity" refers to a substance's affinity for water; the greater the hydrophilicity, the stronger the attraction of the substance to water molecules and the easier it is to be wetted by water.

[0058] In this article, along the direction of liquid flow within the chip, the liquid inlet and outlet of each structure (such as each channel) are referred to as the first end and the second end, respectively.

[0059] Referring to Figure 2, in this text, the trapezoidal shape of a protrusion (such as the first protrusion 132 or the second protrusion 133) means that the width of the protrusion gradually decreases from its first end (i.e., the end adjacent to the first sample dispensing hole) to its second end (i.e., the end adjacent to the sample dispensing channel). For example, referring to Figure 1, the trapezoidal shape of the first protrusion means that the distance between the third side L3 and the fourth side L4 of the first protrusion gradually decreases from the first end to the second end.

[0060] Further, as shown in Figure 1, the protrusion is formed by sequentially connecting a second side L2, a third side L3, a fourth side L4, and a fifth side L5. At least one of the second sides L2 of the protrusion is positioned facing a sample dispensing channel 12, forming a second transition channel II on each side of the sample dispensing inlet of the sample dispensing channel 12. The fifth side L5 is positioned along the edge of the first sample dispensing hole. Preferably, the fifth side L5 is arranged in an arc shape that is the same as or similar to the edge of the first sample dispensing hole.

[0061] The applicant noted that when using microfluidic chips for sample separation, especially with viscous blood samples, blockage often occurs at the inlet of the separation channel. For example, blood samples typically contain fibrin, which can easily clog the separation channel when it clumps together, leading to separation failure or reduced efficiency. To address this, this application proposes a novel microfluidic chip design with anti-clogging capabilities to mitigate or avoid this problem to some extent.

[0062] The chip of this invention includes: a body; a first sample loading port 11, the opening of which is disposed on the upper surface of the body; at least one sample dispensing channel 12 communicating with the first sample loading port, and an anti-clogging region 13 is provided between the sample dispensing channel 12 and the first sample loading port 11; wherein, the anti-clogging region includes:

[0063] A transition section 131 is provided along the edge of the first sample dispensing hole 11 and located on the lower surface of the body. At least two first protrusions 132 are arranged at intervals on the transition section 131 facing the lower surface of the body. A first transition channel I is formed between adjacent first protrusions 132, and a second transition channel II is formed between the first protrusion and the first side of the transition section. Thus, when a sample is added through the first sample dispensing hole, the sample in the first sample dispensing hole can enter the sample dispensing channel through the first transition channel I and the second transition channel II.

[0064] Referring to Figures 1-5, the present invention provides a microfluidic chip with anti-clogging function, comprising:

[0065] Body 10 (also known as chip card or sample card);

[0066] A first sample dispensing port 11, the opening of which is disposed on the upper surface of the body (Figures 8 and 9 show the upper surface of a microfluidic chip); at least one first number of dispensing channels 12 (preferably capillary channels) communicating with the first sample dispensing port, and an anti-clogging region 13 is provided between the dispensing channels 12 and the first sample dispensing port 11; wherein, the anti-clogging region includes:

[0067] A transition section 131 is provided along the edge of the first sample dispensing hole 11 and located on the lower surface of the body. At least two first protrusions 132 are arranged at intervals on the transition section 131 facing the lower surface of the body. A second number of first transition channels I are formed between at least two adjacent first protrusions 132 (the two intersections of two adjacent first protrusions and the edge of the first sample dispensing hole form a first entrance I1, or in other words, a first entrance is formed between the first ends of two adjacent first protrusions). A third number of second transition channels II are formed between the first protrusion and the first side of the transition section (the first side is the side of the transition section that connects to the sample dispensing channel). The sample located in the first sample dispensing hole can enter the sample dispensing channel by passing through the first transition channel I and the second transition channel II in sequence by a driving force (such as capillary action).

[0068] Specifically, in an exemplary embodiment, the anti-clogging region includes a transition zone 131 located on the lower surface of the body, which is formed by raising the transition zone 131 by a first height in a direction away from the lower surface. The transition zone 131 includes a first surface 131a formed by raising the transition zone away from the lower surface, and a second surface 131b connected to the lower surface. The sample inlet of the sample dispensing channel 12 is located on the second surface 131b, while the first surface 131a is a quasi-annular region surrounding at least a portion of the edge of the first sample dispensing hole 11.

[0069] For example, a ring-like shape refers to a geometric pattern formed by a portion of a ring. Of course, in other embodiments, a ring-like shape can also refer to a geometric pattern formed by the arrangement of elliptical rings, as long as its first surface can be distributed along the edge of the first sample application hole to guide the sample to enter the transition zone evenly from all sides. This invention does not limit this.

[0070] Referring to Figure 2, the first end of the first sample dispensing hole 11 is the sample dispensing port, and the second end of the first sample dispensing hole 11 is raised to a first height H1 in a direction away from the lower surface to form a transition zone 131, and the first height H1 can be set at approximately 0.1-0.5 mm. Specifically, the transition zone 131 is set along the edge of the first sample dispensing hole 11 to form a kind of annular region, thereby guiding the liquid to be evenly dispersed from the edge of the first sample dispensing hole into multiple sample dispensing channels.

[0071] At least two first protrusions 132 are arranged at intervals on the first surface 131a, protruding towards the lower surface. A second number of first transition channels I are sequentially formed between adjacent first protrusions 132, and the width of the first transition channel I gradually increases from its first end to its second end (or, the first transition channel I is set to a trumpet shape). A third number of second transition channels II are formed between the at least two first protrusions and the second surface 131b, and at least one of the second transition channels II is adjacent to and connected to the sample inlet.

[0072] Preferably, in some embodiments, the arrangement of the first protrusion satisfies the following rule: the first quantity is less than or equal to the second quantity, and the second quantity is less than the third quantity.

[0073] Furthermore, in some embodiments, the arrangement of the first protrusion also satisfies the following rule: the third quantity is greater than or equal to twice the second quantity.

[0074] This invention provides a two-stage transition channel, which features a triple arrangement in terms of setting order, number, and channel size. Specifically, the invention sequentially sets a slightly larger first transition channel and a second transition channel with a size similar to the sample dispensing channel, with the number of second transition channels exceeding the number of first transition channels. This allows the sample to actively pass through the transition channels under capillary action, enabling the transition and sample dispensing processes to be completed automatically and synchronously. The slightly larger, funnel-shaped first transition channel facilitates rapid sample intake from the first sample feeding hole 11 and guides the sample to disperse further into the second transition channel. The second transition channel, on the one hand, has a size similar to the sample dispensing channel, and on the other hand, is dispersed on both sides of the sample dispensing inlet. This allows the sample to undergo secondary filtration while continuing to enter the sample dispensing channel solely under capillary action, avoiding dependence on external driving forces (such as centrifugal force).

[0075] Furthermore, the above-mentioned distributed two-stage transition channel scheme can not only automatically complete the synchronous operation of transition and sample separation, but also has a certain tolerance for blockage problems. Even if a blood sample with particularly high viscosity causes some channels to be blocked, enough drainage channels (i.e., the pathway formed by at least one first transition channel and at least one second transition channel for connecting the first sample loading port and the sample separation channel) can be formed to ensure the effectiveness of sample separation.

[0076] Furthermore, in some embodiments, referring to Figure 6, the first end 12 of the capillary channel (i.e., the sample dispensing channel 12) is connected to the first sample dispensing port, and the second end of the capillary channel is provided with a flow-limiting region 15. The flow-limiting region 15 includes a flow-limiting channel 15a, the first end of which is connected to the capillary channel, and the width of the flow-limiting channel 15a gradually increases from its first end to its second end. A first through hole 15b is provided at the second end of the flow-limiting channel 15a. The inner diameter of the first through hole 15b is approximately 0.5-1.5 mm.

[0077] Furthermore, the chip also includes a connecting channel 16 disposed in the body, and the connecting channel 16 is connected to the current limiting channel 15a through a first through hole 15b. The width of the connecting channel 16 is greater than the diameter of the first through hole 15b.

[0078] Correspondingly, in some embodiments, as shown in Figures 10 and 11, the chip further includes a patch layer 20, wherein the patch layer 20 is used to seal the channels on the body.

[0079] For example, in some embodiments, the bonding layer 20 is provided with an auxiliary area 21, which includes a first number of first auxiliary areas 211. The first auxiliary area 211 is a first groove adapted to the shape of the sampling channel. When the bonding layer 20 is bonded to the body, the first groove and the sampling channel on the body can be connected to form a complete sampling channel. A first absorbent layer is provided on the groove. The first absorbent layer is made of a hydrophilic material so that the complete sampling channel has a first hydrophilic ability. In this embodiment, a "complete sampling channel" means that the circumferential direction of the sampling channel is tightly connected to the bonding layer through the wall of the body, with two openings formed only at both ends of the channel for liquid inlet and outlet.

[0080] Furthermore, in some embodiments, the auxiliary region 21 further includes a second auxiliary region 212 connected to the first auxiliary region 211. The second auxiliary region has a second groove that is at least adapted to the shape of the second surface. When the adhesive layer 20 is adhered to the body, the second groove, the first surface, and the second surface form a complete transition region. The second groove on the second auxiliary region 212 is provided with a second absorbent layer made of a hydrophilic material to give it hydrophilic properties. Consequently, the sample in the first sample application hole can pass through the first and second transition channels under the guidance of the second absorbent layer.

[0081] Furthermore, in some embodiments, the present invention provides a detection card that cooperates with a sample application card (i.e., the body), and the detection card 30 includes a plurality of reaction chambers 31, as shown in FIG12. A sealing membrane is provided on the opening of the reaction chamber, and a puncture end is provided at the second end of the connecting channel 16, which can puncture the sealing membrane to facilitate the addition of the sample into the reaction chamber.

[0082] Preferably, in order to prevent the liquid sample entering the reaction chamber from flowing back into the sample card, the inner diameter of the reaction chamber is approximately 1.2 mm to 1.4 mm, so that when the test card is placed horizontally or nearly horizontally, the reagent or liquid sample inside will hardly flow when not subjected to external force.

[0083] Typically, the reaction chamber is pre-filled with reagents, including gel and working solution. It is understood that the type of working solution can be flexibly selected according to different detection requirements. The sample application card and the detection card together form the detection assembly.

[0084] In some embodiments, when the detection component is applied to the ABO blood group system, three reaction chambers can be set on the detection card, wherein the working solutions in two reaction chambers include anti-A and anti-B respectively, and the other reaction chamber can serve as a control (i.e., the working solution does not contain antibodies).

[0085] In some embodiments, when the detection component is applied to the Rh system (ISBT 004), six reaction chambers can be set on the detection card, with the working solutions in five of the chambers containing anti-D, anti-C, anti-c, anti-E, and anti-e, respectively, and the remaining chamber serving as a control. Alternatively, when the detection component is applied to the Kell system (ISBT 006), six reaction chambers can also be set on the detection card, with the working solutions in five of the chambers containing anti-D, anti-C, anti-c, anti-E, and anti-e, respectively, and the remaining chamber may additionally contain anti-K (AntiK). Anti-K primarily targets the Kell antigen (K antigen) and is an important antibody for transfusion reactions and neonatal hemolytic disease.

[0086] In some embodiments, when the detection component is applied to the Duffy system (ISBT 008), three reaction chambers can be provided, and the working fluid in two of the reaction chambers respectively includes: anti-Fy a (AntiFy a ) and anti-Fy b (AntiFy b Another reaction chamber can be used as a control.

[0087] In some embodiments, when the detection component is applied to the Kidd system (ISBT 009), three reaction chambers can be provided, and the working fluid in two of the reaction chambers respectively includes: anti-Jk a (AntiJk a ) and anti-Jk b (AntiJk b Another reaction chamber can be used as a control.

[0088] Of course, it is understandable that when the detection component is applied to different types of detection systems, different types of antibodies can be added to the detection card to complete the detection. For example, for the MNS system (ISBT 002), anti-M and anti-N antibodies can be added to the working solutions of the two reaction chambers of the detection card, respectively. As another example, for the Lewis system (ISBT 007), anti-Lewis antibodies can be added to the working solutions of the two reaction chambers of the detection card, respectively. a Anti-Le b For example, for the P1PK system (ISBT 003), anti-P1 can be added to the working fluid in one reaction chamber of the test card. For example, for the Hh system (ISBT 018), anti-H can be added to the working fluid in one reaction chamber. For example, for the Ii system (ISBT 027), anti-I can be added to the working fluid in one reaction chamber.

[0089] In some embodiments, as shown in FIG2, the transition section 131 is a groove provided along the edge of the first sample feeding hole 11. Specifically, the lower surface of the body is recessed inward to form an annular transition section.

[0090] In some embodiments, at least one of the first protrusions corresponds to a sample inlet arrangement of one of the sample dispensing channels, wherein the width of one side of the first protrusion corresponding to the sample inlet (i.e., the second side L2 of the first protrusion 132) is greater than the width of the sample inlet, so as to form two second transition channels II on both sides of the sample inlet by cooperating with the first side. This embodiment employs a multi-transition channel design to ensure that the sample (e.g., blood) passes through at least two transition channels during its entry into the sample dispensing channel from the first sample application port, thereby effectively alleviating the problem of blockage caused by impurities such as fibers in the sample.

[0091] Preferably, in some embodiments, a first protrusion is provided at the sampling inlet of each or multiple sampling channels.

[0092] In some embodiments, the width of the first transition channel I is minimal at its entrance.

[0093] In some embodiments, the width of the entrance of the first transition channel I ranges from 0.05 to 0.25 mm.

[0094] In some embodiments, the width of the first protrusion gradually decreases along the direction from its first end (i.e., the end adjacent to the first sample feeding hole) to its second end.

[0095] In some embodiments, the width of the entrance of the first transition channel is smaller than the channel width of the first transition channel.

[0096] The width of the inlet of the first transition channel I refers to the width of the position where the first transition channel connects to the first sample feeding port. The channel width refers to the width of the first transition channel I at the channel between the inlet and outlet (or, the channel width can also refer to the average width of the first transition channel).

[0097] In some embodiments, the width of the second transition channel II ranges from 0.05 to 0.25 mm.

[0098] In some embodiments, the inlet width of the first transition channel I is greater than the width of the second transition channel II. In this embodiment, the dual transition channel design enables large-size filtration of agglomerates of different sizes (such as fibrin) and at least one small-size filtration. The combination of at least two filtration operations optimizes the filtration effect (preventing excessive fibrin from entering the sample distribution channel and affecting subsequent experimental results), while further preventing blockage of the transition channel inlet.

[0099] In some embodiments, a waste liquid pool 14 is provided at the end of the first sample dispensing port 11 away from the sample dispensing channel. A first transition channel I is also formed between the sidewall of the waste liquid pool 14 and the adjacent first protrusion 132, and a second inlet I2 is formed on the side of the first transition channel I that contacts the first sample dispensing port. In this embodiment, the use of multiple protrusions (such as the first and second protrusions) in a dispersed arrangement along the transition interval can form multiple transition channels, increasing in number, thereby reducing the requirement for centrifugal force in sample filtration while ensuring effective filtration of samples such as blood.

[0100] Furthermore, these numerous and short distributed transition channels can avoid or mitigate clogging problems that may occur during the filtration process.

[0101] In some embodiments, the first protrusion is trapezoidal in design, wherein the first transition channel formed between two adjacent first protrusions gradually increases in size from its transition inlet to its transition outlet. That is, the first transition channel is funnel-shaped. The funnel-shaped transition channel I, which directly communicates with the first sample dispensing port, can reduce the resistance of the transition zone to the sample separation process to a certain extent, and reduce the centrifugal force required for sample separation. For example, even if some impurities enter the first transition channel, the remaining liquid still has a chance to pass through smoothly.

[0102] In some embodiments, when the distance between adjacent first protrusions 132 is greater than a preset distance (for example, in some embodiments, when the distance between adjacent first protrusions is greater than the width of the entrance of a set first transition channel), a second protrusion 133 is also provided between adjacent first protrusions 132, and the third and fourth sides of the second protrusion cooperate with the first protrusions on both sides to form a first supplementary channel and a second supplementary channel.

[0103] In some embodiments, the second protrusion is trapezoidal or triangular in shape, and the width of the first or second supplementary channel gradually increases along its inlet to outlet direction.

[0104] In some embodiments, the widths of the first supplementary channel and the second supplementary channel also gradually increase from their first end (i.e., the end near the first sample loading hole) to their second end.

[0105] In some embodiments, the anti-blocking area is arranged in a ring shape.

[0106] In some embodiments, the height of the first protrusion ranges from 0.05 to 0.20 mm.

[0107] In some embodiments, the number of the first transition channel I and the second transition channel II is greater than the number of the sampling channels.

[0108] In some embodiments, the number of the second transition channels II is greater than twice the number of the sampling channels.

[0109] In some embodiments, the through hole can be a capillary structure, in which case the through hole is equivalent to an extension of the corresponding channel. For example, both the through hole and the sample distribution channel have capillary action, and the inner diameter of the connecting channel is larger than the inner diameter of the through hole, so the liquid will not continue to enter the connecting channel.

[0110] Alternatively, in other embodiments, the through-hole can also be a non-capillary structure, as long as it can serve to limit the flow.

[0111] Obviously, the amount of sample added to each reaction chamber can be the same or different by adjusting the length and / or inner diameter of each sample dispensing channel.

[0112] In some embodiments, the sample dispensing orifice may also adopt a tapered structure that is larger at the top and smaller at the bottom, meaning that the inner diameter of the sample dispensing orifice gradually decreases along the direction from the upper surface to the lower surface of the body. The larger dispensing orifice facilitates sample alignment, while the smaller bottom makes it easier for the sample to contact the dispensing channel, thus speeding up the dispensing process.

[0113] In some embodiments, the two adjacent sides of the first protrusion are connected by an arc structure. Therefore, even when red blood cells in a blood sample are rapidly propelled into the transition area by external forces (such as capillary or centrifugal forces), they are less likely to break.

[0114] In some embodiments, the width of the inlet of the first transition channel I ranges from 0.05 to 0.25 mm, the inner diameter of the through hole is set between approximately 0.5 and 1.5 mm, the width of the sample dispensing channel is between approximately 0.12 and 0.18 mm, and the depth (i.e., the length in the thickness direction H) is between approximately 0.1 and 0.5 mm (wherein, the size of the sample dispensing channel is preferably set to accommodate just about 1 μL of liquid sample).

[0115] The present invention also provides a chip with a reagent replenishment function (see Figures 7-10). For example, the chip in this embodiment further includes:

[0116] The second sample feeding hole 17 has an opening on the upper surface;

[0117] The sample dispensing channel 18 has a first end connected to the second sample dispensing hole 17, and a second end connected to the connecting channel 16 through a second through hole 19; wherein the first through hole 15b is located above the second through hole 19.

[0118] The width of the sample addition channel 18 is greater than the width of the sample separation channel 12, and the width of the connecting channel 16 is less than the width of the connecting channel 16.

[0119] Preferably, the inner diameter of the sample dispensing channel is greater than approximately 0.5 mm.

[0120] In this embodiment, a second sample well is preferably provided, and the second sample well is connected to a communicating channel 16 to facilitate the addition of antibody to a reaction chamber. For example, in some embodiments, the second sample well can be used to add antibody K.

[0121] In this embodiment, the setting of the second sample loading well can improve the flexibility of the detection card in different application scenarios, that is, different antibodies can be added according to the detection needs of different application scenarios or different detection items.

[0122] For example, in some embodiments, the test card operates as follows: First, a second sample (such as antibody K) is added to the second sample well. Then, the test card is centrifuged, typically at a force of 200g for more than 10 seconds. During this time, antibody K enters the connecting channel through the second through-hole and then into the reaction chamber. Subsequently, a first sample (such as a blood sample) is added to the first sample well, and centrifuged again, allowing the blood sample to enter the reaction chamber and react separately with the antibody to obtain the test result.

[0123] Of course, in other embodiments, the present invention utilizes the size difference and the difference in hydrophilicity achieved by the bonding layer to achieve simultaneous addition of the first and second samples and a single centrifugation operation.

[0124] For example, the present invention also provides a microfluidic detection method, comprising the steps of:

[0125] S100, a microfluidic chip as described in any of the above embodiments is provided; wherein, the sample dispensing channel 12 is configured to have a first hydrophilicity, the sample application channel 18 is configured to have a third hydrophilicity, and the third hydrophilicity is greater than or equal to the first hydrophilicity; the width of the sample application channel 18 is greater than the width of the sample dispensing channel 12, and the length of the sample application channel 18 is less than the length of the sample dispensing channel 12;

[0126] In this embodiment, the hydrophilicity can be achieved by means of an adhesive layer. For example, the adhesive layer may also be provided with a third groove that is adapted to the shape of the sample dispensing channel 18, and the third groove is filled with a hydrophilic material to form a third hydrophilic layer.

[0127] S101, a first sample (e.g., blood sample) is added to the first sample well 11, and a second sample (e.g., antibody K) is added to the second sample well 17. The first sample enters the dispensing channel via capillary action, passing through the first transition channel I and the second transition channel II, and stops at the flow-limiting zone 15. The first sample must overcome the first resistance f1 applied by the flow-limiting zone 15 to enter the connecting channel 16. The second sample enters the sample dispensing channel 18 via capillary action and must overcome the second resistance f2 to enter the connecting channel. The second resistance f2 is less than the first resistance f1.

[0128] S102, the chip is centrifuged using a first centrifugal force f3, and the first centrifugal force f3 satisfies a first centrifugation rule: f1-f3>λ1, f3-f2>λ2, where λ1 is a first set value and λ2 is a second set value;

[0129] S103, the chip is further centrifuged using a second centrifugal force f4, and the second centrifugal force f4 satisfies the second centrifugation rule: f4-f1>λ3, where λ3 is a first set value.

[0130] For example, in some embodiments, the first centrifugal force is greater than the second resistance, but less than the first resistance. The second centrifugal force is greater than the first resistance. This allows for the sequential addition of antibodies and liquid samples through a continuous centrifugation process.

[0131] In this embodiment, the first centrifugal force is less than the second centrifugal force. Therefore, after adding the first and second samples, a single continuous centrifugation can be performed, requiring only adjustment of the centrifugal force. This continuous centrifugation operation reduces the complexity of the sample addition process. Furthermore, allowing the antibody to be rapidly mixed into the reagent before adding the liquid sample and reacting with the reagent improves reaction efficiency and thus enhances the reliability of the detection results.

[0132] In some embodiments, the second sample dispensing port 17 includes: a sample dispensing port disposed on the body, and a guide area disposed between the sample dispensing port and the sample dispensing channel, wherein the width of the guide area is smaller than the width of the sample dispensing port, so that the second sample can enter the sample dispensing channel more smoothly.

[0133] In this embodiment, "centrifugal force" refers to the force exerted on the liquid in its flow direction due to centrifugal action during the centrifugation process of the chip.

[0134] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0135] Through the above description of the embodiments, those skilled in the art can clearly understand that the methods of the above embodiments can be implemented by means of software plus necessary general-purpose hardware platforms. Of course, they can also be implemented by hardware, but in many cases the former is a better implementation method. Based on this understanding, some technical solutions of the present invention, or the parts that contribute to the prior art, can also be embodied in the form of software products. This computer software product is stored in a storage medium (such as ROM / RAM, magnetic disk, optical disk) and includes several instructions to cause a computer terminal (which may be a mobile phone, computer, server, or network device, etc.) to execute the methods described in the various embodiments of the present invention.

[0136] The embodiments of the present invention are described above in conjunction with the accompanying drawings, but the present invention is not limited to the above-mentioned specific implementation methods. The above-mentioned specific implementation methods are merely illustrative and not restrictive. Under the guidance of the present invention, ordinary technicians in this field can also make many forms without departing from the scope of protection of the present invention and the claims, all of which are protected by the present invention.

Claims

1. A microfluidic chip, characterized in that, include: ontology; The first sample feeding hole (11) has an opening on the upper surface of the body. A first number of sampling channels (12), each of which is connected to the first sample dispensing hole, and an anti-clogging area (13) is provided between the sampling channel (12) and the first sample dispensing hole (11); wherein, the anti-clogging area includes: A transition section (131) is provided along at least a portion of the edge of the first sample dispensing hole (11) and located on the lower surface of the body. The transition section (131) is formed by raising the height by a first height in a direction away from the lower surface. The transition section (131) includes a first surface (131a) formed by the elevation and a second surface (131b) connected to the lower surface. The sample dispensing inlet of the sample dispensing channel (12) is provided on the second surface (131b), and the first surface (131a) is a quasi-annular region surrounding at least a portion of the edge of the first sample dispensing hole (11). At least two first protrusions (132) are arranged at intervals on the first surface (131a) and protrude toward the lower surface. A second number of first transition channels (I) are formed sequentially between adjacent first protrusions (132) among the at least two first protrusions (132), and the width of the first transition channel (I) gradually increases along the direction from its first end to its second end. The at least two first protrusions respectively form a third number of second transition channels (II) between themselves and the second surface (131b), and at least one second transition channel (II) is adjacent to and connected to the sample dispensing inlet; wherein, the sample located in the first sample feeding hole can enter the sample dispensing channel by passing through the first transition channel (I) and the second transition channel (II) in sequence under the action of driving force; The first protrusion is configured to satisfy the following rule: the first quantity is less than or equal to the second quantity, and the second quantity is less than the third quantity.

2. The microfluidic chip according to claim 1, characterized in that, The third quantity is greater than or equal to twice the second quantity.

3. The microfluidic chip according to claim 1, characterized in that, At least one pair of the second transition channels (II) are respectively disposed on both sides of one of the sample inlets.

4. The microfluidic chip according to claim 1, characterized in that, The height of the first protrusion is 0.05-0.20 mm.

5. The microfluidic chip according to claim 1, characterized in that, A second protrusion (133) is provided between at least two adjacent first protrusions (132), and the third and fourth sides of the second protrusion cooperate with the first protrusions on both sides to form a first supplementary channel and a second supplementary channel.

6. The microfluidic chip according to claim 1, characterized in that, The first entrance of the first transition channel is greater than or equal to the width of the second transition channel, and the width of the second transition channel is the same as or similar to the width of the sampling channel.

7. The microfluidic chip according to claim 1, characterized in that, Also includes: A patch layer (20) is used to seal at least one channel.

8. The microfluidic chip according to claim 7, characterized in that, The second end of the sampling channel (12) is provided with a flow-limiting area (15), the flow-limiting area (15) includes: a flow-limiting channel (15a), the width of the flow-limiting channel (15a) gradually increases along the direction from its first end to its second end, and the second end of the flow-limiting channel (15a) is connected to the connecting channel (16) provided on the body through a first through hole (15b); correspondingly, an auxiliary area (21) is provided on the bonding layer (20), the auxiliary area (21) includes: a first number of first auxiliary areas (211), and the first auxiliary area... The auxiliary area is a first groove adapted to the shape of the sampling channel (12), wherein the first groove is connected to the sampling channel and forms a complete sampling channel, and a first absorbent layer is provided on the first groove. The first absorbent layer is made of a hydrophilic material so that the complete sampling channel has a first hydrophilic ability; the adhesive layer (20) is connected to the flow restriction area and forms a complete flow restriction area so that the complete flow restriction area has a second hydrophilic ability, and the first hydrophilic ability is greater than or equal to the second hydrophilic ability.

9. The microfluidic chip according to claim 8, characterized in that, The bonding layer is a hydrophobic film.

10. The microfluidic chip according to claim 8, characterized in that, Also includes: The second sample feeding hole (17) has an opening on the upper surface; A sample dispensing channel (18) is provided, the first end of which is connected to the second sample dispensing hole (17), and the second end of which is connected to the connecting channel (16) through the second through hole (19); wherein the first through hole (15b) is located above the second through hole (19); The width of the sample addition channel (18) is greater than the width of the sample separation channel (12), and the width of the connecting channel (16) is less than the width of the connecting channel (16).

11. A microfluidic detection method, characterized in that, Including the following steps: S100, providing a microfluidic chip as described in claim 10; wherein the complete sample dispensing channel is configured to have a first hydrophilicity, the sample addition channel is configured to have a third hydrophilicity, and the third hydrophilicity is greater than or equal to the first hydrophilicity; the width of the sample addition channel is greater than the width of the sample dispensing channel, and the length of the sample addition channel is less than the length of the sample dispensing channel; S101, a first sample is added to the first sample feeding hole and a second sample is added to the second sample feeding hole respectively; wherein, the first sample can enter the sample dispensing channel by passing through the first transition channel (I) and the second transition channel (II) in sequence under capillary action, and stop at the flow restriction zone, and the first sample needs to overcome the first resistance f1 applied by the flow restriction zone before it can continue to enter the connecting channel; the second sample can enter the sample feeding channel (18) under capillary action, and needs to overcome the second resistance f2 before it can continue to enter the connecting channel, and the second resistance f2 is less than the first resistance f1; S102, the microfluidic chip is centrifuged using a first centrifugal force f3, and the first centrifugal force f3 satisfies a first centrifugation rule: f1-f3>λ1, f3-f2>λ2, where λ1 is a first set value and λ2 is a second set value; S102, the microfluidic chip is centrifuged using the second centrifugal force f4, and the second centrifugal force f4 satisfies the second centrifugation rule: f4-f1>λ3, where λ3 is a third set value.

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