WO2018184402A1 - 可保留径向高阶模式的封装型光微流微腔生化传感器 - Google Patents

可保留径向高阶模式的封装型光微流微腔生化传感器 Download PDF

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WO2018184402A1
WO2018184402A1 PCT/CN2017/116259 CN2017116259W WO2018184402A1 WO 2018184402 A1 WO2018184402 A1 WO 2018184402A1 CN 2017116259 W CN2017116259 W CN 2017116259W WO 2018184402 A1 WO2018184402 A1 WO 2018184402A1
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microcavity
refractive index
optical
sensor capable
fiber
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French (fr)
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吴翔
汤婷
李子豪
王伯文
孙弘毅
徐雷
刘丽英
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Fudan University
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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01D—MEASURING NOT SPECIALLY ADAPTED FOR A SPECIFIC VARIABLE; ARRANGEMENTS FOR MEASURING TWO OR MORE VARIABLES NOT COVERED IN A SINGLE OTHER SUBCLASS; TARIFF METERING APPARATUS; MEASURING OR TESTING NOT OTHERWISE PROVIDED FOR
    • G01D5/00—Mechanical means for transferring the output of a sensing member; Means for converting the output of a sensing member to another variable where the form or nature of the sensing member does not constrain the means for converting; Transducers not specially adapted for a specific variable
    • G01D5/26—Mechanical means for transferring the output of a sensing member; Means for converting the output of a sensing member to another variable where the form or nature of the sensing member does not constrain the means for converting; Transducers not specially adapted for a specific variable characterised by optical transfer means, i.e. using infrared, visible, or ultraviolet light
    • G01D5/268—Mechanical means for transferring the output of a sensing member; Means for converting the output of a sensing member to another variable where the form or nature of the sensing member does not constrain the means for converting; Transducers not specially adapted for a specific variable characterised by optical transfer means, i.e. using infrared, visible, or ultraviolet light using optical fibres
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
    • G01N21/41—Refractivity; Phase-affecting properties, e.g. optical path length

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  • the invention belongs to the technical field of sensors, and in particular relates to a packaged optical microfluidic microcavity biochemical sensor capable of retaining a radial high-order mode.
  • optical microcavity sensors have been widely used in biochemical sensing due to their ultra-high sensitivity and ultra-low detection limits.
  • An optical microcavity is an optical cavity that is comparable in size to the wavelength of a resonant light. It localizes the light field in the cavity, causing the photon to oscillate multiple times within the cavity, increasing the number of interactions between light and matter. It senses the presence of an analyte by detecting changes in the optical mode within the cavity and can therefore be used to detect the concentration of biomolecules or chemicals.
  • the advantages of the whispering gallery mode microcavity are high Q value, long photon lifetime, and strong interaction of photons with analytes, so that higher detection sensitivity can be obtained in biochemical sensing; in addition, the line of the whispering mode in the microcavity The narrower width facilitates lower detection limits in sensing.
  • the photomicrofluidic microcavity is a type of optical whispering mode microcavity, including a microbubble cavity and a microcavity. They have a unique mid-pass structure that creates a microfluidic channel.
  • the microfluidic channel transports the substance to be tested into the cavity core, which can make the substance to be tested interact with the mode field exposed in the cavity to a greater extent, thereby further improving the sensing performance.
  • photomicrofluidic microcavity biochemical sensors are considered to be effective methods for studying nanometer-scale particles such as nucleic acids, proteins, and viruses.
  • the optical microfluidic microbubble for sensing and the fused cone fiber coupled thereto are exposed to the air, so the sensing process is susceptible to interference from the external environment.
  • the microbubble cavity and the melt cone fiber are connected to the cumbersome five-dimensional adjustment table and cannot be carried around, so they cannot be used in practical biochemical sensing applications.
  • various packaging methods have been proposed. In these packaging methods, the optical microcavity and the coupling fiber are completely encapsulated in a layer of low refractive index polymer, which can overcome the interference of the external environment and be stable and portable.
  • the external environment of the microbubble cavity is air, and the microbubble cavity of the package is wrapped with a layer of low refractive index polymer, so that the relative refractive index of the microbubble cavity and the external environment after encapsulation is reduced, thereby causing light in the microcavity.
  • the critical angle of total reflection of internal propagation increases, eventually causing the original radial high-order mode in the microcavity to disappear after encapsulation.
  • the radial high-order mode in the microfluidic microbubble cavity has higher mode sensitivity because it has more cavity leakage in the cavity and can interact better with the solution material in the cavity. .
  • the object of the present invention is to provide a packaged optical microfluidic microcavity biochemical sensor capable of retaining a radial high-order mode to compensate for the shortcomings of the existing packaged microcavity biochemical sensor, and to maintain a compact structure and be subjected to external conditions.
  • the advantages of low vibration and low detection limit continue to preserve the high-order radial high-order mode in the cavity after packaging.
  • the invention relates to a packaged optical microfluidic microcavity biochemical sensor capable of retaining a radial high-order mode, comprising: a molten cone fiber, an optical microfluidic microbubble cavity, a cover glass, and a UV for encapsulation and fixation.
  • Glue and low refractive index polymers as shown in Figure 1.
  • the photomicrofluidic microcavity is a whispering wall mode microcavity
  • the fused cone fiber is placed on the optical microfluidic microcavity
  • the fused cone optical fiber and the optical microfluidic microcavity are coupled to form a microcavity coupling system, the fusion cone optical fiber and the optical microflow
  • the two ends of the microcavity are respectively fixed by a low refractive index polymer and an ultraviolet glue to fix the relative position of the coupling
  • the cover glass covers the microbubble fiber coupling system
  • the low refractive index polymer and the ultraviolet glue form a barrier after curing
  • the barrier effect of the cover glass forms a confined space centered on the microcavity coupling system, which isolates the influence of water vapor, dust, air pollutants and platform vibration on the coupling system in the external environment.
  • the inner wall of the microfluidic microcavity can be coated with a specific ligand of the analyte to specifically detect the biological sample to be tested.
  • the interval or coupling pitch of the optical microfluidic microcavity and the melt cone optical fiber is between 0-1 ⁇ m.
  • the main component of the melt-cone fiber is silica, and the smallest diameter is 2-5 ⁇ m.
  • the main component of the photomicrofluidic microcavity is silica, having a diameter of 100-500 ⁇ m and a wall thickness of 2-20 ⁇ m.
  • the material at both ends of the fixed melt-cone fiber is a low-refractive-index polymer having a refractive index between 1-1.4 to avoid light leakage.
  • the materials at both ends of the fixed photomicrofluidic microcavity are mainly ultraviolet glue, which is cured by irradiation with an ultraviolet lamp.
  • the principle of the present invention is as follows: First, for the optical microfluidic microcavity sensor of the whispering gallery mode, when the refractive index of the solution in the cavity changes or there is particle adsorption on the inner wall of the cavity, the displacement of the resonant wavelength of the whispering mode is caused, and The partial mode light field is distributed in the cavity core, so the interaction between the light field and the material is strong, and it is easy to obtain higher sensitivity. Secondly, the low refractive index polymer and the ultraviolet glue are respectively used to fix both ends of the melt cone fiber and the optical microfluidic microcavity, thereby fixing the relative positions of the coupling.
  • the cover slip can be used to form a peripherally sealed stable device centered on the microcavity.
  • the encapsulated microcavity environment is still air, so that the radial high-order mode in the microcavity can be preserved.
  • the main component of the melt-cone fiber is silica, with a diameter of 2-5 ⁇ m, which is placed on the micro-fluid microcavity to excite the whispering wall mode in the microcavity;
  • the main component of the photo-microfluidic microcavity is silicon dioxide.
  • the diameter is 100-500 ⁇ m and the wall thickness is 2-20 ⁇ m;
  • the materials at both ends of the fixed fiber are mainly low refractive index polymers, the refractive index is between 1-1.4, which can avoid light leakage;
  • the materials at both ends of the fixed optical microfluidic microcavity Mainly UV glue, cured by UV light.
  • the transmission spectrum of one or more optical whispering gallery modes can be obtained from the output of the cone fiber, as shown in FIG. Radial high-order modes in photomicrofluidic microcavities tend to have higher sensitivity.
  • the sensing of the sample to be tested can be performed by monitoring the displacement of the single mode resonant wavelength or the relative amount of change of the multiple mode resonant wavelengths.
  • the device of the invention senses by monitoring the displacement of the optical mode resonant wavelength, and has the advantages of compact structure, small size, strong anti-interference, good stability, simple manufacturing process and low cost.
  • the device can retain the radial high-order mode in the coupled microcavity, so it has a very low detection limit and ultra-high sensitivity, and has a good application prospect in the field of biochemical sensing.
  • FIG. 1 is a schematic diagram showing the structure of a packaged optical microfluidic microcavity biochemical sensor that can retain a radial high-order mode.
  • 2(a), (b), and (c) are pattern transmission spectra of the microcavity before, after, and after 24 hours.
  • the resonant wavelengths are all around 1550 nm, and the air is in the microcavity.
  • Figure 3 is a measurement of the microcavity quality factor.
  • Figure 4 is a graph showing the difference in the resonance modes caused by the solution of different concentrations of small molecule compounds (Biotin D-Biotin). Among them, (a) is 0.84 pm, and (b) is 1.77 pm.
  • Bioenzymes were specifically identified using the encapsulated photomicrofluidic biochemical sensor.
  • the measurement wavelength used is around 780 nm
  • the diameter of the cone fiber is 3 ⁇ m, and it is placed on the microfluidic microcavity, that is, the coupling pitch is 0 ⁇ m;
  • the microcavity is a microfluidic microbubble microcavity, as shown in Fig. 1, the microcavity has a diameter of about 350 ⁇ m and a wall thickness of 4 ⁇ m;
  • the low refractive index encapsulant is MY133 polymer with a refractive index of 1.33. Since this packaging technology keeps the microcavity still in the air, the radial high-order mode in the microcavity can be preserved. As shown in Fig. 2, the microcavity sensor after 24 hours of packaging is still clear due to the stable anti-interference of the package structure. Mode transmission spectrum;
  • the small molecule compound (Biotin D-Biotin) was subjected to a sensing test using the encapsulated microcavity prepared above.
  • the surface treatment was first performed to adhere a specific antibody to the inner surface of the microcavity, and then the Biotin solution of 50 fg/mL and 100 fg/mL was respectively introduced to measure the displacement of the resonance mode.
  • two concentrations of Biotin solution were introduced into the microcavity causing a red shift of the resonant mode of 0.84 pm and 1.77 pm, respectively.
  • the biochemical sensor can also be used to detect other biomolecule antigens such as the p24 protein antigen carried on the surface of HIV virus, and can also achieve extremely low detection limits;
  • the self-reference technology such as mode splitting or mode difference can be used to suppress the noise of the signal, eliminate the influence of the external environment, and further improve the detection limit of the sensor.

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Abstract

一种可保留径向高阶模式的封装型光微流微腔生物化学传感器，包括：一段熔锥光纤，一个光微流微腔，一块盖玻片，以及封装固定用紫外胶和低折射率的聚合物；光微流微腔主要成分为二氧化硅，直径为100-500微米，壁厚为2-20微米；熔锥光纤主要成分为二氧化硅，直径为2-5微米，用于激发微腔中的回音壁模式；低折射率聚合物的折射率在1-1.4之间，用于固定熔锥光纤两端以及防止光泄露。通过监测光学模式谐振波长的位移情况来进行传感，具有结构紧凑、尺寸小、抗干扰性强、稳定性好、制作工艺简单以及成本低等优点。此外，能够保留耦合微腔中的径向高阶模式，因此具有极低探测极限以及超高灵敏度，在生物化学传感领域有良好的应用前景。

Description

可保留径向高阶模式的封装型光微流微腔生化传感器 技术领域
本发明属传感器技术领域，具体涉及一种可保留径向高阶模式的封装型光微流微腔生物化学传感器。
 背景技术
近些年，光学微腔传感器因其超高灵敏度和超低探测极限在生物化学传感领域应用广泛。光学微腔是指尺寸与谐振光波长相比拟的光学谐振腔，它将光场局域在腔里，令光子在腔内多次震荡，增加了光与物质相互作用的次数。它通过探测腔内光学模式的变化来感知分析物的存在，因此可以用于探测生物分子或者化学物质的浓度。回音壁模式微腔的优点是Q值高，光子寿命长，光子与分析物的相互作用强，从而在生物化学传感中可以获得更高的探测灵敏度；此外，微腔中回音壁模式的线宽较窄，有利于在传感中获得更低的探测极限。
光微流微腔是光学回音壁模式微腔中一种类型，包括微泡腔和微管腔。它们拥有独特的中通结构，可以形成微流通道。微流通道将待测物质运输至腔芯中，可以使待测物质与露在腔内的模场更大程度地相互作用，从而进一步提高传感性能。目前，光微流微腔生物化学传感器被认为是对核酸、蛋白质、病毒等尺度在纳米量级的颗粒进行研究的有效手段。
一般地，用于传感的光微流微泡腔和与其耦合的熔锥光纤均暴露在空气中，因此传感过程易受外部环境的干扰。此外，微泡腔和熔锥光纤均与笨重的五维调整台相连，不可随身携带，因而无法真正应用于实际的生物化学传感应用中。为了解决上述问题，多种封装方式被提出。在这些封装方式中，光学微腔和耦合光纤均被完全地包裹在一层低折射率的聚合物中，可以克服外界环境的干扰并且稳定便携。然而，封装之前微泡腔外界环境为空气，封装之微泡腔外包裹着一层低折射率聚合物，因此封装后微泡腔与外部环境的相对折射率减小，从而引起光在微腔内传播的全反射临界角度增大，最终导致微腔内原本的径向高阶模式在封装之后消失。实验和计算表明，光微流微泡腔中的径向高阶模式由于其模场泄露在腔内比例更多，可以和腔内的溶液物质更好地互相作用，因此具有更高的模式灵敏度。然而传统的全包裹式封装方法会使微腔中的径向高阶模式消失，因此我们提出一种新的封装型光微流微腔传感器，既稳固便携又能保留腔内径向高阶模式从而获得较高的传感灵敏度。
 技术问题
本发明的目的在于提出一种可保留径向高阶模式的封装型光微流微腔生物化学传感器，以弥补现有的封装型微腔生物化学传感器存在的不足，既保持结构紧凑、受外界震动影响小以及探测极限低等优点，还在封装后继续保留腔内具有高灵敏度的径向高阶模式。
 技术解决方案
本发明提出的可保留径向高阶模式的封装型光微流微腔生物化学传感器，包括：一段熔锥光纤，一个光微流微泡腔，一块盖玻片，以及用于封装固定的紫外胶和低折射率聚合物，如图1所示。
其中，光微流微腔为回音壁模式微腔，熔锥光纤搭靠在光微流微腔上，熔锥光纤与光微流微腔耦合组成微腔耦合系统，熔锥光纤与光微流微腔的两端分别由低折射率聚合物和紫外胶固定，从而固定二者耦合的相对位置；盖玻片覆盖在微泡光纤耦合系统上方；低折射率聚合物和紫外胶固化后形成屏障，加上盖玻片的阻隔作用，形成以微腔耦合系统为中心的密闭空间，隔绝了外界环境中水汽、灰尘、空气中的污染物以及平台震动对耦合系统的影响。
本发明中，微流微腔内壁可以涂覆待测物的特异性配体，从而对被测生物样品进行特异性探测。
本发明中，光微流微腔与熔锥光纤的间隔或者耦合间距为0-1 μm 之间。
本发明中，熔锥光纤主要成分为二氧化硅，直径最细处为2-5μm。
本发明中，光微流微腔的主要成分为二氧化硅，直径为100-500μm，壁厚为2-20μm。
本发明中，固定熔锥光纤两端的材料为低折射率聚合物，其折射率在1-1.4之间，可以避免光泄露。
本发明中，固定光微流微腔两端的材料主要为紫外胶，通过紫外灯照射固化。
本发明原理如下：首先，对于回音壁模式的光微流微腔传感器而言，当其腔内溶液折射率发生改变或腔内壁上有颗粒吸附时，会引起回音壁模式谐振波长的位移，并且其部分模式光场会分布于腔芯中，因此光场与物质的相互作用很强，易于获得较高的灵敏度。其次，低折射率聚合物和紫外胶分别用于固定熔锥光纤与光微流微腔的两端，从而固定二者耦合的相对位置。加上盖玻片可以形成以微腔为中心的四周密封型稳固器件。封装后的微腔外界环境仍然是空气，从而可以保留微腔中的径向高阶模式。熔锥光纤主要成分为二氧化硅，直径为2-5μm，搭靠在光微流微腔上，用以激发出微腔中的回音壁模式；光微流微腔的主要成分为二氧化硅，直径为100-500μm，壁厚为2-20μm；固定光纤两端的材料主要为低折射率聚合物，折射率在1-1.4之间，可以避免光泄露；固定光微流微腔两端的材料主要为紫外胶，通过紫外灯照射固化。从熔锥光纤的输出端可以得到一个或多个光学回音壁模式的透射谱，如图2所示。光微流微腔中的径向高阶模式，往往有较高的灵敏度。通过监测单个模式谐振波长的位移量或者多个模式谐振波长的相对变化量就可以对待测样品进行传感探测。
 有益效果
本发明器件通过监测光学模式谐振波长的位移情况来进行传感，具有结构紧凑、尺寸小、抗干扰性强、稳定性好、制作工艺简单以及成本低等优点。此外，本器件能够保留耦合微腔中的径向高阶模式，因此具有极低探测极限以及超高灵敏度，在生物化学传感领域有良好的应用前景。
 附图说明
图1是可保留径向高阶模式的封装型光微流微腔生物化学传感器结构示意图。
图2（a）、（b）、（c）分别为微腔在封装前、封装后以及24小时后的模式透射谱图。谐振波长均在1550 nm附近，微腔中均为空气。
图3是对微腔品质因子测量结果。
图4是测量不同浓度的小分子化合物（生物素D-Biotin）溶液引起谐振模式不同的位移量。其中，（a）为0.84pm，和（b）为1.77 pm。
 本发明的实施方式
下面通过具体实施例进一步描述本发明：
实施例：利用该封装型光微流生物化学传感器对生物分子进行特异性识别。
1、采用的测量波长在780nm附近；
2、熔锥光纤直径为3μm，搭靠在微流微腔上，即其耦合间距为0μm；
3、微腔为光微流微泡型微腔，如图1所示，微腔直径约为350μm ，壁厚为4μm；
4、低折射率封装材料采用MY133聚合物，其折射率为1.33。由于此封装技术保持微腔仍处于空气中，因此可以保留微腔中的径向高阶模式，如图2所示，由于封装结构稳固抗干扰，封装24小时后的微腔传感器仍有清晰的模式透射谱；
5、用于传感的薄壁微腔中的典型谐振模式及其品质因子如图3所示。若采用厚壁微腔进行封装，品质因子约为10 7；
6、采用上述制备的封装型微腔对小分子化合物（生物素D-Biotin）进行传感测试。为了特异性结合Biotin，首先进行表面处理，使得微腔内表面黏附一层特异性抗体，再分别通入50fg/mL和100 fg/mL的Biotin溶液，测量谐振模式的位移情况。如图4所示，两种浓度的Biotin溶液通入微腔中分别引起谐振模式0.84pm和1.77 pm的红移。该生物化学传感器亦可用于探测其他的生物分子抗原如HIV病毒表面携带的p24蛋白质抗原，同样能达到极低的探测极限；
7、实验中，可以采用模式劈裂或者模式差分等自参考技术对信号的噪声进行抑制，消除外界环境的影响，进一步提高传感器的探测极限。

Claims (6)

  1. 可保留径向高阶模式的封装型光微流微腔生物化学传感器，其特征在于，包括：一段熔锥光纤，一个光微流微腔，一块盖玻片，以及用于封装固定的紫外胶和低折射率聚合物；其中，光微流微腔为回音壁模式微腔，熔锥光纤搭靠在光微流微腔上，熔锥光纤与光微流微腔耦合组成微腔耦合系统，熔锥光纤与光微流微腔的两端分别由低折射率聚合物和紫外胶固定，从而固定二者耦合的相对位置；盖玻片覆盖在微泡光纤耦合系统上方；低折射率聚合物和紫外胶固化后形成屏障，加上盖玻片的阻隔作用，形成以微腔耦合系统为中心的密闭空间。
  2. 根据权利要求1所述的可保留径向高阶模式的封装型光微流微腔生物化学传感器，其特征在于，光微流微腔与熔锥光纤的间隔或者耦合间距为0-1 μm之间。
  3. 根据权利要求1所述的可保留径向高阶模式的封装型光微流微腔生物化学传感器，其特征在于，熔锥光纤主要成分为二氧化硅，直径最细处为2-5μm。
  4. 根据权利要求1所述的可保留径向高阶模式的封装型光微流微腔生物化学传感器，其特征在于，光微流微腔的主要成分为二氧化硅，直径为100-500μm，壁厚为2-20μm。
  5. 根据权利要求1所述的可保留径向高阶模式的封装型光微流微腔生物化学传感器，其特征在于，固定熔锥光纤两端的材料为低折射率聚合物，其折射率在1-1.4之间。
  6. 根据权利要求1所述的可保留径向高阶模式的封装型光微流微腔生物化学传感器，其特征在于，固定光微流微腔两端的材料主要为紫外胶，通过紫外灯照射固化。
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