WO2022253145A1 - 一种血液检测微流控芯片 - Google Patents
一种血液检测微流控芯片 Download PDFInfo
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- WO2022253145A1 WO2022253145A1 PCT/CN2022/095792 CN2022095792W WO2022253145A1 WO 2022253145 A1 WO2022253145 A1 WO 2022253145A1 CN 2022095792 W CN2022095792 W CN 2022095792W WO 2022253145 A1 WO2022253145 A1 WO 2022253145A1
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- pool
- waste liquid
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- plasma
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Images
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502753—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/02—Adapting objects or devices to another
- B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
- B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
- B01L2200/10—Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N2015/0019—Means for transferring or separating particles prior to analysis, e.g. hoppers or particle conveyors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N2015/0092—Monitoring flocculation or agglomeration
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/01—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials specially adapted for biological cells, e.g. blood cells
- G01N2015/018—Platelets
Definitions
- the invention relates to the technical fields of molecular detection and microfluidic chips, and more specifically relates to a blood detection microfluidic chip.
- the coagulation system of the human body is a rather complex system, including coagulation factors and anticoagulation factors, fibrinolytic system and antifibrinolytic system, etc.
- Coagulation detection is of great significance for disease prevention, disease treatment and blood transfusion therapy.
- problems in blood sample storage, sample extraction and pretreatment, detection efficiency and detection timeliness in coagulation detection are:
- Blood coagulation testing requires cumbersome sample extraction and pretreatment processes, and each step needs to be equipped with fixed instruments and consumables, which are prone to problems such as operational errors and sample contamination, which affect the reliability of testing;
- the traditional blood coagulation test separates the blood sample from the test, each step takes a certain amount of time, the test efficiency is low, and timely or batch testing cannot be achieved;
- the present invention provides a microfluidic chip for blood detection to at least solve one of the problems raised in the background technology section above.
- a blood detection microfluidic chip including a substrate and a cover plate, the cover plate and the substrate are sealed and fitted to form a chip body; the chip body is provided with several separation detection units, and several separation detection units The units are distributed radially with the center of the chip body as the origin;
- the separation and detection unit includes a whole blood sampling pool, a plasma separation pool, an erythrocyte sedimentation pool, a plasma storage pool, a quantitative pool, a pre-stage primary waste liquid pool, a pre-stage secondary waste liquid pool, The rear primary waste liquid pool, the detection reagent sampling pool, the detection pool, and the whole blood sampling hole set on the cover plate, the whole blood sampling pool vent hole, the plasma storage pool vent hole, and the rear primary stage Vent hole of waste liquid pool, detection reagent injection hole;
- the whole blood sampling pool is set near the center of the chip body, and the whole blood sampling pool, the plasma separation pool, the erythrocyte sedimentation pool, the quantification pool and the detection pool are separated along the The direction of the center of the chip body is arranged sequentially; the plasma storage pool is located on one side of the red blood cell sedimentation pool; the front primary waste liquid pool and the rear primary waste liquid pool are respectively located side; the detection reagent injection pool is located on the side of the rear primary waste liquid pool;
- the whole blood sampling pool communicates with the plasma separation pool through a first separation channel, the plasma separation pool communicates with the red blood cell sedimentation pool through a second separation channel, and the plasma separation pool communicates with the red blood cell sedimentation pool
- the pools are also connected through communication channels;
- the plasma separation pool is communicated with the plasma storage pool through a siphon channel, and the plasma storage pool is connected with the pre-stage primary waste liquid pool, the post-position primary waste liquid pool, and the quantitative pool through a liquid separation pipe. Connected; the front primary waste liquid pool communicates with the front secondary waste liquid pool through the first circulation channel;
- the second flow channel connected to the quantitative pool merges with the third flow channel connected to the detection reagent injection pool, and the fourth flow channel connected to the detection pool, and a microfluidic valve is set at the intersection;
- the whole blood sampling hole and the air hole of the whole blood sampling pool correspond to and communicate with the whole blood sampling pool;
- the detection reagent injection holes correspond to and communicate with the detection reagent sampling pool;
- the vent hole of the rear primary waste liquid pool corresponds to and communicates with the post primary waste liquid pool;
- the vent hole of the plasma storage pool corresponds to and communicates with the plasma storage pool.
- the whole blood sampling pool, the plasma separation pool, the erythrocyte sedimentation pool, the plasma storage pool, the quantitative pool, the pre- The primary waste liquid pool, the pre-stage secondary waste liquid pool, the rear primary waste liquid pool, the detection reagent injection pool, the detection pool, the first separation channel, the second The separation channel, the communication channel, the siphon channel, the liquid separation channel, the first flow channel, the second flow channel, the third flow channel, and the fourth flow channel are formed by etching Or by cutting, a groove structure is formed on the side of the substrate facing the cover plate.
- the whole blood sampling pool is in the shape of a hook, and the whole blood sampling hole corresponds to and communicates with the longer end of the whole blood sampling pool , the air hole of the whole blood sampling pool corresponds to and communicates with the shorter end of the whole blood sampling pool.
- a cover plate fixing hole is provided at the center of the cover plate, and a substrate fixing hole is provided at the center of the substrate; the shape of the cover plate fixing hole and the substrate fixing hole are The same and overlapped arrangement form a chip fixing hole penetrating through the chip body.
- the material of the cover plate and the substrate is one of silicon wafer, quartz, glass or polymer compound.
- the polymer compound is polymethacrylate or polystyrene or cycloolefin copolymer or polycarbonate.
- the present invention discloses a microfluidic chip for blood detection, which has both separation and detection functions, and can realize the integration of plasma/serum separation, reagent injection, reaction and detection. , to solve the problem that blood coagulation measurement cannot meet the growing demand for real-time detection.
- the blood detection microfluidic chip disclosed by the present invention does not require any pre-treatment of blood samples and chips, and does not need to embed other drugs or touch devices, only by centrifugation
- the blood coagulation detection can be completed by action driving, and the chip can be used in many fields such as blood detection, immune detection, in vitro diagnosis and medical cosmetology.
- FIG. 1 is the structural representation of the present invention
- Figure 2 is a schematic diagram of the structure of the separation and detection unit.
- the embodiment of the present invention discloses a blood detection microfluidic chip, which includes a substrate and a cover plate, and the cover plate and the substrate are sealed and fitted to form a chip body 1; the chip body 1 is provided with several separation detection units 2, and several separation detection units 2 are arranged on the chip body 1.
- the detection units 2 are radially distributed with the center of the chip body 1 as the origin;
- the separation and detection unit 2 includes a whole blood sampling pool 3, a plasma separation pool 4, an erythrocyte sedimentation pool 5, a plasma storage pool 6, a quantification pool 7, a pre-stage primary waste liquid pool 8, and a pre-stage secondary waste liquid pool arranged on the substrate.
- the whole blood sampling pool 3 is set close to the center of the chip body 1, and the whole blood sampling pool 3, the plasma separation pool 4, the red blood cell sedimentation pool 5, the quantification pool 7 and the detection pool 12 are arranged in sequence along the direction away from the center of the chip body 1.
- the plasma storage pool 6 is located on one side of the red blood cell sedimentation pool 5; the front primary waste liquid pool 8 and the rear primary waste liquid pool 10 are respectively located on both sides of the quantitative pool 7; the detection reagent injection pool 11 is located at the rear primary level One side of the waste liquid pool 10;
- the whole blood sampling pool 3 communicates with the plasma separation pool 4 through the first separation channel 18, the plasma separation pool 4 communicates with the red blood cell sedimentation pool 5 through the second separation channel 19, and the plasma separation pool 4 communicates with the red blood cell sedimentation pool 5. It is also communicated through the communication channel 20 to achieve the effect of fluid communication;
- the plasma separation pool 4 communicates with the plasma storage pool 6 through the siphon channel 21, which can achieve a reasonable transfer effect; the plasma storage pool 6 is connected to the front primary waste liquid pool 8, the rear primary waste liquid pool 10 and the Quantitative pool 7 is connected to ensure quantification and transfer; the front primary waste liquid pool 8 communicates with the front secondary waste liquid pool 9 through the first circulation channel 23;
- the second flow channel 24 connected to the quantitative pool 7 merges with the third flow channel 25 connected to the detection reagent injection pool 11 and the fourth flow channel 26 connected to the detection pool 12, and a microfluidic valve 27 is set at the intersection to ensure that the detection pool 12 Mix evenly and prevent the backflow of the test solution;
- the whole blood sampling hole 13 and the air hole 14 of the whole blood sampling pool correspond to and communicate with the whole blood sampling pool 3, which can realize fluid circulation and separation;
- the detection reagent sampling hole 17 corresponds to and communicates with the detection reagent sampling pool 11;
- the vent hole 16 of the rear primary waste liquid pool corresponds to and communicates with the post primary waste liquid pool 10 ;
- the vent hole 15 of the plasma storage pool corresponds to and communicates with the plasma storage pool 6 .
- whole blood sample pool 3 plasma separation pool 4, erythrocyte sedimentation pool 5, plasma storage pool 6, quantification pool 7, pre-stage primary waste pool 8, pre-stage secondary waste pool 9 , Post primary waste liquid pool 10, detection reagent sampling pool 11, detection pool 12, first separation channel 18, second separation channel 19, communication channel 20, siphon channel 21, liquid separation channel 22, first circulation channel 23.
- the second flow channel 24, the third flow channel 25, and the fourth flow channel 26 are groove structures formed on the side of the substrate facing the cover plate by etching or cutting.
- the whole blood sampling pool 3 is in the shape of a hook, and the whole blood sampling hole 13 corresponds to and communicates with the longer end of the whole blood sampling pool 3, and the air hole 14 of the whole blood sampling pool is connected to the whole blood sampling pool.
- the shorter end of the blood sample pool 3 corresponds to and communicates with.
- a cover plate fixing hole is provided in the center of the cover plate, and a substrate fixing hole is provided in the center of the substrate; the cover plate fixing hole and the substrate fixing hole have the same shape and are overlapped to form a chip fixing hole 28 penetrating through the chip body 1.
- the chip fixing hole 28 it is used for cooperating with the centrifugal rotating device to install, so that the chip can be rotated, and different plasma samples can be obtained by adjusting the centrifugal force and centrifugation time.
- the material of the cover plate and the substrate is one of silicon wafer, quartz, glass or polymer compound.
- the polymer compound is polymethacrylate or polystyrene or cycloolefin copolymer or polycarbonate.
- multiple detection reagent injection pools 11 can be provided, which are respectively communicated with the detection pools 12 .
- the whole blood is separated into two parts: plasma and red blood cell sedimentation, the plasma enters the plasma separation pool 4, and the red blood cell sedimentation enters the red blood cell sedimentation pool 5;
- the plasma is filled with the front primary waste liquid pool 8 and the quantitative pool 7, and excess plasma enters the rear primary waste liquid pool 10, and the quantitative pool 7 plays a quantitative role;
- the sample in the pre-stage primary waste liquid pool 8 enters the pre-stage secondary waste liquid pool 9, and at the same time, the excess plasma in the plasma separation pool 4, siphon channel 21 and plasma storage pool 6 enters In the pre-secondary waste liquid pool 9, it will not affect the quantitative pool 7; the detection reagent is added to the detection reagent injection pool 11, and the plasma in the quantitative pool 7 is mixed with the detection reagent at 2000-3500r/min 3-10s and enters the detection Pool 12, followed by detection.
- PRP platelet-rich plasma can be obtained at 3000-3500r/min 60s clockwise
- PPP platelet-poor plasma can be obtained at 4500-5500r/min120-180s;
- the first separation pipe and the second separation pipe have a liquid separation function driven by centrifugal force; the quantitative pool 7 has a quantitative function; and the microfluidic valve 27 has a function of increasing fluid resistance.
- Platelet aggregation test the PRP plasma and PPP plasma in the embodiment can be used for platelet aggregation test, the separated PRP and PPP are centrifuged into the detection hole, and the platelet aggregation analyzer automatically adds the platelet aggregation inducer, generally more ADP (diphosphate Adenosine), and one of ADR (adrenaline), COL (collagen), ARA (arachidonic acid), and RIS (ristocetin) can also be used as the inducer.
- ADP diphosphate Adenosine
- ADR adrenaline
- COL collagen
- ARA arachidonic acid
- RIS ristocetin
- the microfluidic chip blood separation and detection chip in the present invention integrates separation and detection, which can realize one-step separation and detection, and the chip does not require any pretreatment, embedding other drugs or touch devices, and only relies on centrifugation
- the blood coagulation detection can be completed by the function driving, and the operation is simple, the efficiency is high, and the reliability is strong.
- the innovative design of the chip in the present invention includes a siphon channel 21, a waste liquid pool, a separation channel, a quantitative pool 7 and a flow valve, which play the roles of effective transfer, detection and quantification, and prevention of back sucking, increasing detection accuracy.
- the present invention solves the cumbersome sample extraction and pretreatment process in traditional detection, and effectively avoids reliability problems caused by operational errors and sample contamination.
- the chip in the present invention can prepare different plasma samples, such as PRP, PPP, etc., by adjusting the centrifugation rate, which is simple, fast and practical.
- the present invention solves the problem of limited utilization of traditional blood sample extraction, saves sample consumption, and improves sample utilization.
- each embodiment in this specification is described in a progressive manner, each embodiment focuses on the difference from other embodiments, and the same and similar parts of each embodiment can be referred to each other.
- the description is relatively simple, and for the related information, please refer to the description of the method part.
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Abstract
一种血液检测微流控芯片,包括基板和盖板,盖板与基板之间密封配合形成芯片本体(1);芯片本体(1)上设置有若干个分离检测单元(2),若干个分离检测单元(2)以芯片本体(1)的圆心为原点,呈放射状分布;分离检测单元(2)包括设置在基板上的全血进样池(3)、血浆分离池(4)、红细胞沉淀池(5)、血浆储存池(6)、定量池(7)、前置一级废液池(8)、前置二级废液池(9)、后置一级废液池(10)、检测试剂进样池(11)、检测池(12),以及设置在盖板上的全血进样孔(13)、全血进样池透气孔(14)、血浆储存池透气孔(15)、后置一级废液池透气孔(16)、检测试剂进样孔(17)。血液检测微流控芯片同时具有分离和检测功能,可实现血浆/血清分离、试剂注入、反应与检测一体化,解决凝血测量检测无法满足日益增长的即时检测需求问题。
Description
本发明涉及分子检测和微流控芯片技术领域,更具体的说是涉及一种血液检测微流控芯片。
人体的凝血系统是一个颇为复杂的系统,其中包括凝血因子和抗凝血因子,纤溶系统和抗纤溶系统等。
凝血检测对于疾病预防、疾病治疗和输血治疗具有重要意义。然而目前,凝血检测仍存在血样储存、样本提取与前处理、检测效率及检测及时性等方面的问题,其主要局限性在于:
1、血液样本不宜长时间储存,随着时间的推移易发生检测指标变化和降解的风险;
2、凝血检测需要繁琐的样本提取和前处理过程,且每一步骤均需搭配固定的仪器与耗材,易存在操作误差、样本污染等问题,影响检测可靠性;
3、传统的凝血检测血样分离与检测分开,每步处理均需一定时间,检测效率低,且不能实现及时或批量检测;
4、传统检测血样需求量大,但血样采集量有限。
因此,如何提供一种集分离检测为一体的血液检测微流控芯片是本领域技术人员亟需解决的问题。
发明内容
有鉴于此,本发明提供了一种血液检测微流控芯片,以至少解决上述背景技术部分所提出的问题之一。
为了实现上述方案,本发明采用以下技术方案:
一种血液检测微流控芯片,包括基板和盖板,所述盖板与所述基板之间密封配合形成芯片本体;所述芯片本体上设置有若干个分离检测单元,若干个所述分离检测单元以所述芯片本体的圆心为原点,呈放射状分布;
所述分离检测单元包括设置在所述基板上的全血进样池、血浆分离池、红细胞沉淀池、血浆储存池、定量池、前置一级废液池、前置二级废液池、后置一级废液池、检测试剂进样池、检测池,以及设置在所述盖板上的全血进样孔、全血进样池透气孔、血浆储存池透气孔、后置一级废液池透气孔、检测试剂进样孔;
所述全血进样池设置在靠近所述芯片本体的圆心处,并且所述全血进样池、所述血浆分离池、所述红细胞沉淀池、所述定量池和所述检测池沿远离所述芯片本体圆心方向依次设置;所述血浆储存池位于所述红细胞沉淀池一侧;所述前置一级废液池和所述后置一级废液池分别位于所述定量池的两侧;所述检测试剂进样池位于所述后置一级废液池一侧;
所述全血进样池与所述血浆分离池之间通过第一分离通道连通,所述血浆分离池与所述红细胞沉淀池通过第二分离通道连通,并且所述血浆分离池与所述红细胞沉淀池之间还通过连通通道连通;
所述血浆分离池通过虹吸通道与所述血浆储存池连通,所述血浆储存池通过分液管道与所述前置一级废液池、所述后置一级废液池和所述定量池连通;所述前置一级废液池通过第一流通通道与前置二级废液池连通;
所述定量池连接的第二流通通道与所述检测试剂进样池连接的第三流通通、所述检测池连接的第四流通通道交汇,并在交汇处设置微流体阀门;
所述全血进样孔和所述全血进样池透气孔与所述全血进样池对应且连通;所述检测试剂进样孔与所述检测试剂进样池对应且连通;所述后置一级废液池透气孔与所述后置一级废液池对应且连通;所述血浆储存池透气孔与所述血浆储存池对应且连通。
优选的,在上述一种血液检测微流控芯片中,所述全血进样池、所述血浆分离池、所述红细胞沉淀池、所述血浆储存池、所述定量池、所述前置一级废液池、所述前置二级废液池、所述后置一级废液池、所述检测试剂进样 池、所述检测池、所述第一分离通道、所述第二分离通道、所述连通通道、所述虹吸通道、所述分液管道、所述第一流通通道、所述第二流通通道、所述第三流通通道、所述第四流通通道为通过刻蚀或者切割方式,在所述基板朝向所述盖板一面形成的凹槽结构。
优选的,在上述一种血液检测微流控芯片中,所述全血进样池呈弯钩状,并且所述全血进样孔与所述全血进样池较长的一端对应且连通,所述全血进样池透气孔与所述全血进样池较短的一端对应且连通。
优选的,在上述一种血液检测微流控芯片中,所述盖板中心设置有盖板固定孔,所述基板中心设置有基板固定孔;所述盖板固定孔与所述基板固定孔形状相同且重合设置,形成贯穿所述芯片本体的芯片固定孔。
优选的,在上述一种血液检测微流控芯片中,所述盖板和所述基板的材质为硅片、石英、玻璃或高分子化合物中的一种。
优选的,在上述一种血液检测微流控芯片中,所述高分子化合物为聚甲基丙烯酸酯或聚苯乙烯或环烯烃共聚物或聚碳酸酯。
经由上述的技术方案可知,与现有技术相比,本发明公开提供了一种血液检测微流控芯片,同时具有分离和检测功能,可实现血浆/血清分离、试剂注入、反应与检测一体化,解决凝血测量无法满足日益增长的即时检测需求问题,本发明公开提供的血液检测微流控芯片,无需对血样和芯片进行任何的前期处理,并且无需包埋其他药物或触动装置,仅靠离心作用驱动即可完成凝血检测,该芯片可用于血液检测、免疫检测、体外诊断和医疗美容等多个领域。
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据提供的附图获得其他的附图。
图1附图为本发明的结构示意图;
图2附图为分离检测单元的结构示意图。
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
本发明实施例公开了一种血液检测微流控芯片,包括基板和盖板,盖板与基板之间密封配合形成芯片本体1;芯片本体1上设置有若干个分离检测单元2,若干个分离检测单元2以芯片本体1的圆心为原点,呈放射状分布;
分离检测单元2包括设置在基板上的全血进样池3、血浆分离池4、红细胞沉淀池5、血浆储存池6、定量池7、前置一级废液池8、前置二级废液池9、后置一级废液池10、检测试剂进样池11、检测池12,以及设置在盖板上的全血进样孔13、全血进样池透气孔14、血浆储存池透气孔15、后置一级废液池透气孔16、检测试剂进样孔17;
全血进样池3设置在靠近芯片本体1的圆心处,并且全血进样池3、血浆分离池4、红细胞沉淀池5、定量池7和检测池12沿远离芯片本体1圆心方向依次设置;血浆储存池6位于红细胞沉淀池5一侧;前置一级废液池8和后置一级废液池10分别位于定量池7的两侧;检测试剂进样池11位于后置一级废液池10一侧;
全血进样池3与血浆分离池4之间通过第一分离通道18连通,血浆分离池4与红细胞沉淀池5通过第二分离通道19连通,并且血浆分离池4与红细胞沉淀池5之间还通过连通通道20连通,可达到流体连通的效果;
血浆分离池4通过虹吸通道21与血浆储存池6连通,可达到合理转移的效果;血浆储存池6通过分液管道22与前置一级废液池8、后置一级废液池10和定量池7连通,可保证定量和转移;前置一级废液池8通过第一流通通道23与前置二级废液池9连通;
定量池7连接的第二流通通道24与检测试剂进样池11连接的第三流通通25、检测池12连接的第四流通通道26交汇,并在交汇处设置微流体阀门27,保证检测池12混合均匀并阻止检测液回流;
全血进样孔13和全血进样池透气孔14与全血进样池3对应且连通,可实现流体流通与分离;检测试剂进样孔17与检测试剂进样池11对应且连通;后置一级废液池透气孔16与后置一级废液池10对应且连通;血浆储存池透气孔15与血浆储存池6对应且连通。
为了进一步优化上述技术方案,全血进样池3、血浆分离池4、红细胞沉淀池5、血浆储存池6、定量池7、前置一级废液池8、前置二级废液池9、后置一级废液池10、检测试剂进样池11、检测池12、第一分离通道18、第二分离通道19、连通通道20、虹吸通道21、分液管道22、第一流通通道23、第二流通通道24、第三流通通25道、第四流通通道26为通过刻蚀或者切割方式,在基板朝向盖板一面形成的凹槽结构。
为了进一步优化上述技术方案,全血进样池3呈弯钩状,并且全血进样孔13与全血进样池3较长的一端对应且连通,全血进样池透气孔14与全血进样池3较短的一端对应且连通。
为了进一步优化上述技术方案,盖板中心设置有盖板固定孔,基板中心设置有基板固定孔;盖板固定孔与基板固定孔形状相同且重合设置,形成贯穿芯片本体1的芯片固定孔28,通过芯片固定孔28,用于与离心旋转设备配合安装,使该芯片进行旋转,可通过调节离心力大小和离心时间,得到不同血浆样品。
为了进一步优化上述技术方案,盖板和基板的材质为硅片、石英、玻璃或高分子化合物中的一种。
为了进一步优化上述技术方案,高分子化合物为聚甲基丙烯酸酯或聚苯乙烯或环烯烃共聚物或聚碳酸酯。
为了进一步优化上述技术方案,根据需要,检测试剂进样池11可以设置有多个,分别与检测池12连通。
检测方法:
将全血加入全血进样池3中,顺时针3000-5500r/min 60-180s,全血分离成为血浆和红细胞沉淀两部分,血浆进入血浆分离池4,红细胞沉淀进入红细胞沉淀池5;
随后逆时针500-1000r/min 30-60s,血浆充满前置一级废液池8和定量池7,多余血浆进入后置一级废液池10,定量池7起到定量作用;
3000-5500r/min 5-30s,前置一级废液池8中样品进入到前置二级废液池9中,同时血浆分离池4、虹吸通道21和血浆储存池6中多余的血浆进入前置二级废液池9中,不会对定量池7造成影响;将检测试剂加入检测试剂进样池11,2000-3500r/min 3-10s定量池7中血浆与检测试剂混合并进入检测池12中,随后检测。
当血浆和红细胞沉淀时,可通过调节离心力大小,得到不同血浆样品,例如顺时针3000-3500r/min 60s可得到PRP富血小板血浆,4500-5500r/min120-180s可得到PPP贫血小板血浆;
第一分离管道和第二分离管道具有在离心驱动下分液作用;定量池7具有定量作用;微流体阀门27具有增大流体阻力的作用。
血小板聚集测试,实施例中的PRP血浆和PPP血浆可用于血小板聚集测试,分离好的PRP和PPP离心到检测孔中,血小板聚集仪全自动加入血小板聚集诱导剂,一般用ADP较多(二磷酸腺苷),诱导剂也可以使用ADR(肾上腺素)、COL(胶原)、ARA(花生四烯酸)、RIS(瑞斯托霉素)中的一种。
按诱导剂:血浆体积比1:5-1:20的比例加入诱导剂,优选的是1:10,加入到试剂孔。离心3000r/min 3s,震荡混匀。在血小板聚集仪上进行检测。记录透光率-时间变化数据,并计算出血小板最大聚集率。对所测定的数据进行处理都属于成熟的技术内容,在此不再进行详细描述。
本发明优越性如下:
1.本发明中的微流控芯片血液分离和检测芯片芯片集分离与检测为一体,可实现一步法分离与检测,且芯片无需任何前处理、无需包埋其他药物或触动装置、仅靠离心作用驱动即可完成凝血检测,操作简单、效率高、可靠性强。
2.本发明中芯片设计创新的包含了虹吸通道21、废液池、分离通道、定量池7与流通阀,起到有效转移、检测定量、防止倒吸的作用,增加检测精准度。
3.本发明解决了传统检测中繁琐的样本提取和前处理过程,有效地避免了操作误差、样本污染带来的可靠性问题。
4.本发明中芯片可通过调节离心速率来制备不同的血浆样品,例如PRP、PPP等,简单、快捷、实用。
5.本发明中解决了传统血样提取利用有限的问题,节约样本用量,提高了样本利用率。
本说明书中各个实施例采用递进的方式描述,每个实施例重点说明的都是与其他实施例的不同之处,各个实施例之间相同相似部分互相参见即可。对于实施例公开的装置而言,由于其与实施例公开的方法相对应,所以描述的比较简单,相关之处参见方法部分说明即可。
对所公开的实施例的上述说明,使本领域专业技术人员能够实现或使用本发明。对这些实施例的多种修改对本领域的专业技术人员来说将是显而易见的,本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下,在其它实施例中实现。因此,本发明将不会被限制于本文所示的这些实施例,而是要符合与本文所公开的原理和新颖特点相一致的最宽的范围。
Claims (6)
- 一种血液检测微流控芯片,其特征在于,包括基板和盖板,所述盖板与所述基板之间密封配合形成芯片本体;所述芯片本体上设置有若干分离检测单元,若干个所述分离检测单元以所述芯片本体的圆心为原点,呈放射状分布;所述分离检测单元包括设置在所述基板上的全血进样池、血浆分离池、红细胞沉淀池、血浆储存池、定量池、前置一级废液池、前置二级废液池、后置一级废液池、检测试剂进样池、检测池,以及设置在所述盖板上的全血进样孔、全血进样池透气孔、血浆储存池透气孔、后置一级废液池透气孔、检测试剂进样孔;所述全血进样池设置在靠近所述芯片本体的圆心处,并且所述全血进样池、所述血浆分离池、所述红细胞沉淀池、所述定量池和所述检测池沿远离所述芯片本体圆心方向依次设置;所述血浆储存池位于所述红细胞沉淀池一侧;所述前置一级废液池和所述后置一级废液池分别位于所述定量池的两侧;所述检测试剂进样池位于所述后置一级废液池一侧;所述全血进样池与所述血浆分离池之间通过第一分离通道连通,所述血浆分离池与所述红细胞沉淀池通过第二分离通道连通,并且所述血浆分离池与所述红细胞沉淀池之间还通过连通通道连通;所述血浆分离池通过虹吸通道与所述血浆储存池连通,所述血浆储存池通过分液管道与所述前置一级废液池、所述后置一级废液池和所述定量池连通;所述前置一级废液池通过第一流通通道与前置二级废液池连通;所述定量池连接的第二流通通道与所述检测试剂进样池连接的第三流通通、所述检测池连接的第四流通通道交汇,并在交汇处设置微流体阀门;所述全血进样孔和所述全血进样池透气孔与所述全血进样池对应且连通;所述检测试剂进样孔与所述检测试剂进样池对应且连通;所述后置一级废液池透气孔与所述后置一级废液池对应且连通;所述血浆储存池透气孔与所述血浆储存池对应且连通。
- 根据权利要求1所述的一种血液检测微流控芯片,其特征在于,所述全血进样池、所述血浆分离池、所述红细胞沉淀池、所述血浆储存池、所述定量池、所述前置一级废液池、所述前置二级废液池、所述后置一级废液池、所述检测试剂进样池、所述检测池、所述第一分离通道、所述第二分离通道、 所述连通通道、所述虹吸通道、所述分液管道、所述第一流通通道、所述第二流通通道、所述第三流通通道、所述第四流通通道为通过刻蚀或者切割方式,在所述基板朝向所述盖板一面形成的凹槽结构。
- 根据权利要求1所述的一种血液检测微流控芯片,其特征在于,所述全血进样池呈弯钩状,并且所述全血进样孔与所述全血进样池较长的一端对应且连通,所述全血进样池透气孔与所述全血进样池较短的一端对应且连通。
- 根据权利要求1所述的一种血液检测微流控芯片,其特征在于,所述盖板中心设置有盖板固定孔,所述基板中心设置有基板固定孔;所述盖板固定孔与所述基板固定孔形状相同且重合设置,形成贯穿所述芯片本体的芯片固定孔。
- 根据权利要求1所述的一种血液检测微流控芯片,其特征在于,所述盖板和所述基板的材质为硅片、石英、玻璃、高分子化合物中的一种。
- 根据权利要求5所述的一种血液检测微流控芯片,其特征在于,所述高分子化合物为聚甲基丙烯酸酯或聚苯乙烯或环烯烃共聚物或聚碳酸酯。
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CN116298347A (zh) * | 2023-05-12 | 2023-06-23 | 北京未神生物科技有限公司 | 一种离心式微流控的全自动血型检测卡及其检测方法 |
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