WO2020119282A1 - 一种双聚焦微流体芯片 - Google Patents

一种双聚焦微流体芯片 Download PDF

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WO2020119282A1
WO2020119282A1 PCT/CN2019/113186 CN2019113186W WO2020119282A1 WO 2020119282 A1 WO2020119282 A1 WO 2020119282A1 CN 2019113186 W CN2019113186 W CN 2019113186W WO 2020119282 A1 WO2020119282 A1 WO 2020119282A1
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dual
microfluidic chip
flow channel
focus
discrete phase
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French (fr)
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于赐龙
钱翔
赵艺
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Shenzhen University
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Shenzhen University
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    • 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
    • H—ELECTRICITY
    • H01—ELECTRIC ELEMENTS
    • H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
    • H01J49/00—Particle spectrometers or separator tubes
    • H01J49/02—Details
    • H01J49/04—Arrangements for introducing or extracting samples to be analysed, e.g. vacuum locks; Arrangements for external adjustment of electron- or ion-optical components
    • H01J49/0431—Arrangements for introducing or extracting samples to be analysed, e.g. vacuum locks; Arrangements for external adjustment of electron- or ion-optical components for liquid samples
    • H01J49/0445—Arrangements for introducing or extracting samples to be analysed, e.g. vacuum locks; Arrangements for external adjustment of electron- or ion-optical components for liquid samples with means for introducing as a spray, a jet or an aerosol
    • H—ELECTRICITY
    • H01—ELECTRIC ELEMENTS
    • H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
    • H01J49/00—Particle spectrometers or separator tubes
    • H01J49/02—Details
    • H01J49/10—Ion sources; Ion guns
    • H01J49/16—Ion sources; Ion guns using surface ionisation, e.g. field-, thermionic- or photo-emission
    • H01J49/165—Electrospray ionisation
    • 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
    • B01L2200/06—Fluid handling related problems
    • B01L2200/0631—Purification arrangements, e.g. solid phase extraction [SPE]

Definitions

  • the invention relates to the field of electrospray mass spectrometry detection, in particular to a dual-focus microfluidic chip.
  • Viscous liquid samples (such as cell suspensions) occupy an important position in the fields of life sciences and industry. Mass spectrometry analysis has become an important method for the molecular level characterization of viscous liquid samples and the measurement of their components.
  • the microfluidic chip has the advantages of small size, multi-functional integration, and low sample consumption. It can perform extraction, separation, and desalting of complex samples and other pretreatment operations, making its combination with mass spectrometry a research hotspot.
  • the sample needs to be introduced into the macro-structure ion source to achieve sample ionization, which is inconvenient to operate, and cannot achieve high-throughput detection, low consumption, and high sensitivity The need for rapid testing.
  • Direct ionization technology generally uses conductive tips (such as probe electrospray PESI) or high pressure gas flow (such as electrospray extraction ionization EESI) to achieve direct ionization of viscous liquid samples under the condition of charged droplet extraction, thereby Reduce the ion suppression effect caused by traditional ionization methods (such as ESI, APCI, etc.), and eliminate the influence of the matrix on the mass spectrometer.
  • conductive tips such as probe electrospray PESI
  • high pressure gas flow such as electrospray extraction ionization EESI
  • EESI electrospray extraction ionization
  • the object of the present invention is to provide a dual-focus microfluidic chip, which aims to solve the problem that the existing microfluidic chip cannot yet directly ionize viscous liquid samples.
  • a dual-focus microfluidic chip including:
  • the discrete phase flow channel is used for guiding the discrete phase.
  • the discrete phase flow channel is provided with a first shrink hole, one end of the discrete phase flow channel is provided with a discrete phase inlet, and the other end is provided with a second shrink hole;
  • the diameter of the first shrinkage hole and the diameter of the second shrinkage hole are both smaller than the width of the discrete phase flow channel;
  • Two first branch channels for guiding the first continuous phase, and the two first branch channels converge at the first shrinkage hole;
  • Two second branch channels are used for guiding the second continuous phase, and the two second branch channels converge at the second shrinkage hole.
  • a plurality of first concave cavities are provided at the junction of the discrete phase inlet and the discrete phase flow channel.
  • two ends of the first branch channels away from the first shrinkage hole converge to form a first dry channel, and the end of the first dry channel is provided with a first continuous phase inlet .
  • a plurality of second cavities are provided at the junction of the first continuous phase inlet and the first dry channel.
  • two ends of the second branch channels away from the second converging hole converge to form a second dry channel, and a second continuous phase inlet is provided at the end of the second dry channel .
  • the dual-focus microfluidic chip wherein the depth of the first branch flow channel is greater than the depth of the discrete phase flow channel;
  • the depth of the second branch flow channel is greater than the depth of the discrete phase flow channel.
  • the dual-focus microfluidic chip is made of PDMS.
  • the discrete phase is a viscous liquid
  • the first continuous phase is an extractant
  • the second continuous phase is a gas
  • the viscous liquid is cell suspension, crude oil or blood.
  • the present invention provides a dual-focus microfluidic chip as described above.
  • the present invention realizes the droplet-liquid-liquid series extraction of viscous samples through secondary three-dimensional fluid focusing, making the viscous samples direct Spray ionization greatly simplifies the pretreatment process of viscous samples. Moreover, the extraction and ionization of smaller-sized droplets in three-dimensional space is conducive to improving the ionization efficiency.
  • FIG. 1 is a structural diagram of an embodiment of a dual-focus microfluidic chip of the present invention.
  • FIG. 2 is a partially enlarged view of A in FIG. 1.
  • FIG. 3 is a structural diagram of another embodiment of a dual-focus microfluidic chip of the present invention.
  • the present invention provides a dual-focus microfluidic chip.
  • the present invention will be described in further detail below. It should be understood that the specific embodiments described herein are only used to explain the present invention, and are not intended to limit the present invention.
  • a preferred embodiment of a dual-focus microfluidic chip provided by the present invention includes a discrete phase flow channel 1 for guiding a discrete phase and two first continuous phases for guiding The first branch channel 21 and two second branch channels 31 for guiding the second continuous phase.
  • the discrete phase flow channel 1 is provided with a first shrinkage hole 11 with an aperture of R1, one end of the discrete phase flow channel 1 is provided with a discrete phase inlet 12, and the other end is provided with a second shrinkage hole 13 with an aperture of R2;
  • R1 and R2 are both Less than the width R0 of the discrete phase flow channel 1, for example, R0 can be set to 100um, R1 and R2 can be set to 50um,; two first branch channels 21 converge at the first shrinkage hole 11; two second branch channels 31 Converge at the second shrink hole 13.
  • the discrete phase is a viscous liquid
  • the first continuous phase is an extractant
  • the second continuous phase is a gas
  • the extractant passes through two first branch channels Focusing with the viscous liquid at the first shrinkage hole for the first time, the viscous liquid is squeezed into a fine stream or a single droplet, producing micron-sized droplets, and the extractant realizes the first droplet extraction; the gas passes through the two The two branch channels and the micron-level droplets are sprayed in the second focusing at the second shrinkage hole.
  • the sample droplets are sprayed under the three-dimensional package of the extractant, which belongs to three-dimensional space liquid-liquid extraction and energy charge.
  • a mass spectrometer can then be used to analyze the atomized droplets.
  • the invention adopts secondary three-dimensional fluid focusing, introduces fluid shear force, and forms a stretching flow, which helps to separate the viscous sample into nano droplets; the extraction and ionization of smaller-sized droplets in three-dimensional space is conducive to improving ions ⁇ Efficiency.
  • the invention can directly use the microchip-mass spectrometry to spray and ionize the viscous sample on the microfluidic chip. It should be emphasized that if there is only one focus, then the spray ionization of the viscous liquid can not be achieved at all. Instead, using two focus, the first focus can cause the microscopic droplets of the viscous sample, and the extractant droplets After the extraction and the second focusing, the gas can realize the direct spray ionization of the viscous sample.
  • the depth of the first branch channel 21 and the depth of the second branch channel 31 are greater than the depth of the discrete phase channel 1, for example, the first branch channel 21 and the second branch channel 31 are set to 300um, and the discrete phase channel 1 is set It is 30um, which is conducive to the discrete phase flowing out of the discrete phase flow channel (205) and suspended in the continuous phase, completely wrapped by the continuous phase, without contacting the wall surface of the flow channel, forming a stretch flow form, completing the separation of the discrete phase and injection.
  • a first dry channel 2 may be provided at an end of the two first branch channels 21 away from the first constriction hole 11, as shown in FIG. 3, a first continuous phase is provided at the end of the first dry channel 2 In the inlet 22, the first continuous phase enters from the first continuous phase inlet 22 and branches to the two first branch channels 21 through the first main flow channel 2.
  • a plurality of second cavities 23 may also be provided at the junction of the first continuous phase inlet 22 and the first main flow channel 2.
  • a plurality of first concave cavities 33 may be provided at the junction of the discrete phase inlet 12 and the discrete phase flow channel 1, and both of the concave cavities may be regular array units to prevent clogging.
  • the discrete phase flow channel 1 and the first branch flow channel 21 may be arranged in a circuitous shape, such as a zigzag structure.
  • a second dry channel 3 may be provided at an end of the two second branch channels 31 away from the second constriction hole 13, as shown in FIG. 3, a second continuous phase inlet is provided at the end of the second dry channel 3 32.
  • the second continuous phase enters from the second continuous phase inlet 32 and branches through the second main flow channel 3 to the two second branch channels 31.
  • the dual-focus microfluidic chip of the present invention can be made of quartz, glass and other materials, preferably polydimethylsiloxane (PDMS), which is easy to process and has low cost.
  • PDMS polydimethylsiloxane
  • the existing multi-layer soft lithography process can be used, and multiple mask plates are used.
  • multiple steps of glue rejection, exposure, and development are carried out in a single silicon wafer
  • the flow channel structure with different depths is formed on the top, and the precise flow channel pattern production is realized through the mask plate.
  • the process is very mature, and the present invention will not repeat them.
  • the upper and lower halves can be made first, and then assembled and assembled to form each flow channel.
  • the present invention provides a dual-focus microfluidic chip.
  • the present invention introduces fluid shearing force through secondary three-dimensional fluid focusing to form a tensile flow, which helps to separate viscous samples into nano droplets;
  • the extraction and ionization of smaller size droplets in three-dimensional space is beneficial to improve the ionization efficiency.
  • the direct ionization performance of viscous samples on the microfluidic chip can be tested. Under the condition of low sample consumption, high-throughput real-time analysis of viscous liquids can be achieved to enable molecular-level analysis Cell biochemical information transformation and regulation becomes possible.

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Abstract

一种双聚焦微流体芯片，包括：离散相流道（1），用于导流离散相，离散相流道（1）上设置有第一缩孔（11），离散相流道（1）的一端设置有离散相入口（12），另一端设置有第二缩孔（13）；第一缩孔（11）的孔径和第二缩孔（13）的孔径均小于离散相流道（1）的宽度；两个第一支流道（21），用于导流第一连续相，两个第一支流道（21）汇聚于第一缩孔（11）处；两个第二支流道（31），用于导流第二连续相，两个第二支流道（31）汇聚于第二缩孔（13）处。

Description

一种双聚焦微流体芯片 技术领域
本发明涉及电喷雾质谱检测领域，尤其涉及一种双聚焦微流体芯片。
背景技术
黏性液态样品(例如细胞悬浊液)在生命科学及工业等领域占有重要位置,质谱分析成为黏性液态样品分子水平表征及其中衡量组分检测的重要方法。
微流控芯片具有尺寸小、多功能集成、样品消耗低等优势，可以对复杂样品进行萃取、分离、除盐等前处理操作，使得其与质谱仪的联用成为研究热点。但是，在微流控芯片上完成细胞培养、裂解、化学反应等操作后，需要将样品引入宏观结构离子源实现样品离子化，操作不便，且无法实现高通量检测、低消耗量以及高灵敏度的快速检测需求。
直接离子化技术一般都是通过导电尖端(如探针电喷雾PESI)或者高压气流(如电喷雾萃取电离EESI)，在带电液滴萃取的条件下实现对黏性液态样品的直接离子化，从而降低传统离子化方法(如ESI、APCI等)带来的离子抑制效应，消除基质对质谱仪器的影响。然而，黏性流体分子间吸引力较大，具有极端复杂基质，现有的微流控芯片缺少导电尖端，致使电场引力和库伦斥力不足以直接离子化黏性液态样品。
因此，现有技术还有待于改进和发展。
发明内容
鉴于上述现有技术的不足，本发明的目的在于提供一种双聚焦微流体芯片，旨在解决现有的微流控芯片还不能够实现对黏性液态样品直接离子化的问题。
本发明的技术方案如下：
一种双聚焦微流体芯片，包括：
离散相流道，用于导流离散相，所述离散相流道上设置有第一缩孔，所述离散相流道的一端设置有离散相入口，另一端设置有第二缩孔；所述第一缩孔的孔径和所述第二缩孔的孔径均小于所述离散相流道的宽度；
两个第一支流道，用于导流第一连续相，两个所述第一支流道汇聚于所述第一缩孔处；
两个第二支流道，用于导流第二连续相，两个所述第二支流道汇聚于所述第二缩孔处。
所述的双聚焦微流体芯片，其中，所述离散相入口与所述离散相流道的衔接处设置有若 干第一凹腔。
所述的双聚焦微流体芯片，其中，两个所述第一支流道远离所述第一缩孔的一端汇聚形成第一干流道，所述第一干流道的末端设置有第一连续相入口。
所述的双聚焦微流体芯片，其中，所述第一连续相入口与所述第一干流道的衔接处设置有若干第二凹腔。
所述的双聚焦微流体芯片，其中，两个所述第二支流道远离所述第二缩孔的一端汇聚形成第二干流道，所述第二干流道的末端设置有第二连续相入口。
所述的双聚焦微流体芯片，其中，所述第一支流道的深度大于所述离散相流道的深度；
和/或所述第二支流道的深度大于所述离散相流道的深度。
所述的双聚焦微流体芯片，其中，所述双聚焦微流体芯片采用PDMS制作而成。
所述的双聚焦微流体芯片，其中，所述离散相为黏性液体，所述第一连续相为萃取剂，所述第二连续相为气体。
所述的双聚焦微流体芯片，其中，所述黏性液体为细胞悬浊液、原油或血液。
有益效果：本发明提供了一种如上所述的所述的双聚焦微流体芯片，本发明通过二次三维流体聚焦，实现对黏性样品的液滴-液液串联萃取，使得黏性样品直接喷雾离子化，极大简化了黏性样品的预处理流程。而且，对更小尺寸液滴在三维空间萃取和离子化，有利于提高离子化效率。
附图说明
图1为本发明的一种双聚焦微流体芯片的实施例结构图。
图2为图1中A的局部放大图。
图3为本发明的另一种双聚焦微流体芯片的实施例结构图。
具体实施方式
本发明提供了一种双聚焦微流体芯片，为使本发明的目的、技术方案及效果更加清楚、明确，以下对本发明进一步详细说明。应当理解，此处所描述的具体实施例仅仅用以解释本发明，并不用于限定本发明。
本发明提供的一种双聚焦微流体芯片的较佳实施例，如图1和图2所示，包括用于导流离散相的离散相流道1、两个用于导流第一连续相的第一支流道21以及两个用于导流第二连续相的第二支流道31。离散相流道1上设置有第一缩孔11，孔径为R1，离散相流道1的一端设置有离散相入口12，另一端设置有第二缩孔13，孔径为R2；R1和R2均小于离散相流 道1的宽度R0，例如，R0可以设置为100um，R1和R2可以设置为50um，；两个第一支流道21汇聚于第一缩孔11处；两个第二支流道31汇聚于第二缩孔13处。
以黏性液体如细胞悬浊液、原油或血液的质谱测试为例，离散相为黏性液体，第一连续相为萃取剂，第二连续相为气体，萃取剂通过两个第一支流道与黏性液体在第一缩孔处第一次聚焦，黏性液体被挤压成细流或单个液滴，产生微米级液滴，萃取剂实现第一次液滴萃取；气体通过两个第二支流道与微米级液滴在第二缩孔处第二次聚焦喷雾，在第二次气动辅助聚焦中，样品液滴在萃取剂三维包裹下喷出，属于三维空间液液萃取和能荷转移，然后可以采用质谱仪对雾化液滴进行样品分析。本发明通过二次三维流体聚焦，引入流体剪切力，形成拉伸流，有助于黏性样品分离成纳米液滴；对更小尺寸液滴在三维空间萃取和离子化，有利于提高离子化效率。
本发明通过微芯片-质谱连用，可在微流控芯片上将黏性样品直接喷雾离子化。需要强调的是，如果只有一次聚焦，那么完全没法实现黏性液体的喷雾离子化，而采用两次聚焦，第一次聚焦，可以使黏性样品产生微小液滴，同时萃取剂进行液滴萃取，再经过第二次聚焦，气体就可以实现黏性样品的直接喷雾离子化了。
在低样品消耗量的条件下，实现对黏性液体的高通量实时分析，使在分子层面上解析细胞的生化信息转化和调控成为可能。
优选的，第一支流道21的深度和第二支流道31的深度大于离散相流道1的深度，例如，第一支流道21和第二支流道31设置为300um，离散相流道1设置为30um，从而有利于离散相从离散相流道(205)流出后悬浮于连续相中，完全被连续相包裹，而不与流道壁面接触，形成拉伸流形式，完成离散相的分离与喷射。
优选的，本发明可以在两个第一支流道21远离第一缩孔11的一端设置第一干流道2，如图3所示，所述第一干流道2的末端设置有第一连续相入口22，第一连续相从第一连续相入口22进入并经第一干流道2分流至两个第一支流道21。还可以在第一连续相入口22与第一干流道2的衔接处设置若干第二凹腔23。本发明还可以在离散相入口12与离散相流道1的衔接处设置若干第一凹腔33，两个凹腔均可以是规律的阵列单元，用于防止堵塞。为防止流体回流，离散相流道1和第一支流道21可设置为迂回形，比如Z字型结构。
具体的，本发明还可以在两个第二支流道31远离第二缩孔13的一端设置第二干流道3，如图3所示，第二干流道3的末端设置有第二连续相入口32。第二连续相从第二连续相入口32进入并经第二干流道3分流至两个第二支流道31。
本发明的双聚焦微流体芯片可采用石英、玻璃等材料制作，优选采用聚二甲基硅氧烷 (polydimethylsiloxane，PDMS)，该材料易于加工，成本低。
关于本发明的双聚焦微流体芯片的制作，可采用现有的多层软光刻加工工艺，使用多块掩膜版，制作过程中进行多次甩胶、曝光、显影步骤，在一块硅片上形成深浅不同的流道结构，通过掩膜版来实现精确的流道图形制作，该工艺非常成熟，本发明不做赘述。具体可以先制作上、下半片，然后再进行贴合组装形成各流道。
综上所述，本发明提供了一种双聚焦微流体芯片，本发明通过二次三维流体聚焦，引入流体剪切力，形成拉伸流，有助于黏性样品分离成纳米液滴；对更小尺寸液滴在三维空间萃取和离子化，有利于提高离子化效率。通过微芯片-质谱连用，可测试微流控芯片上黏性样品的直接离子化性能，在低样品消耗量的条件下，实现对黏性液体的高通量实时分析，使在分子层面上解析细胞的生化信息转化和调控成为可能。
应当理解的是，本发明的应用不限于上述的举例，对本领域普通技术人员来说，可以根据上述说明加以改进或变换，所有这些改进和变换都应属于本发明所附权利要求的保护范围。

Claims (9)

  1. 一种双聚焦微流体芯片，其特征在于，包括：
    离散相流道，用于导流离散相，所述离散相流道上设置有第一缩孔，所述离散相流道的一端设置有离散相入口，另一端设置有第二缩孔；所述第一缩孔的孔径和所述第二缩孔的孔径均小于所述离散相流道的宽度；
    两个第一支流道，用于导流第一连续相，两个所述第一支流道汇聚于所述第一缩孔处；
    两个第二支流道，用于导流第二连续相，两个所述第二支流道汇聚于所述第二缩孔处。
  2. 根据权利要求1所述的双聚焦微流体芯片，其特征在于，所述离散相入口与所述离散相流道的衔接处设置有若干第一凹腔。
  3. 根据权利要求1所述的双聚焦微流体芯片，其特征在于，两个所述第一支流道远离所述第一缩孔的一端汇聚形成第一干流道，所述第一干流道的末端设置有第一连续相入口。
  4. 根据权利要求3所述的双聚焦微流体芯片，其特征在于，所述第一连续相入口与所述第一干流道的衔接处设置有若干第二凹腔。
  5. 根据权利要求1所述的双聚焦微流体芯片，其特征在于，两个所述第二支流道远离所述第二缩孔的一端汇聚形成第二干流道，所述第二干流道的末端设置有第二连续相入口。
  6. 根据权利要求1-5任一所述的双聚焦微流体芯片，其特征在于，所述第一支流道的深度大于所述离散相流道的深度；
    和/或所述第二支流道的深度大于所述离散相流道的深度。
  7. 根据权利要求1-5任一所述的双聚焦微流体芯片，其特征在于，所述双聚焦微流体芯片采用PDMS制作而成。
  8. 根据权利要求1-5任一所述的双聚焦微流体芯片，其特征在于，所述离散相为黏性液体，所述第一连续相为萃取剂，所述第二连续相为气体。
  9. 根据权利要求8所述的双聚焦微流体芯片，其特征在于，所述黏性液体为细胞悬浊液、原油或血液。
PCT/CN2019/113186 2018-12-12 2019-10-25 一种双聚焦微流体芯片 Ceased WO2020119282A1 (zh)

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