CN106483099A - Optical biosensor based on tunable filter and Michelson's interferometer cascade - Google Patents
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Abstract
Description
技术领域technical field
本发明涉及一种光学传感器，尤其涉及一种基于可调滤波器和迈克尔逊干涉仪级联的光学生物传感器。The invention relates to an optical sensor, in particular to an optical biosensor based on cascaded cascaded filters and Michelson interferometers.
背景技术Background technique
生物传感器在生物检测，化学分析和环境监测领域有十分重要的应用。低成本、高灵敏度和实时在线测量是生物传感的几个技术难点。目前，大部分的生物传感器都将生物浓度的变化转换为电信号进行检测，检测时间较长，不能实时在线测量。无标记光学生物传感器是唯一能够直接检测生物分子反应的仪器，可以快速、实时地监控反应过程，具有灵敏度高、抗电磁干扰能力强等一系列优势。目前，基于光学的生物传感技术大量涌现，例如：荧光光谱技术、拉曼散射光谱技术、吸收光谱技术以及基于倏逝波的折射率探测技术。然而，目前大部分商品化的光学生物传感的缺点在于器件体积大，价格昂贵，所需样品量也大。Biosensors have very important applications in the fields of biological detection, chemical analysis and environmental monitoring. Low cost, high sensitivity and real-time online measurement are several technical difficulties of biosensing. At present, most biosensors convert the change of biological concentration into electrical signals for detection, which takes a long time to detect and cannot be measured online in real time. Label-free optical biosensor is the only instrument that can directly detect the reaction of biomolecules. It can monitor the reaction process quickly and in real time, and has a series of advantages such as high sensitivity and strong anti-electromagnetic interference ability. At present, a large number of optical-based biosensing technologies are emerging, such as: fluorescence spectroscopy, Raman scattering spectroscopy, absorption spectroscopy, and refractive index detection technology based on evanescent waves. However, most of the current commercial optical biosensing devices are bulky, expensive, and require a large amount of samples.
本发明提供的基于可调周期性滤波器和迈克尔逊干涉仪级联的无标记光学生物传感器由可调周期性滤波器与和迈克尔逊干涉仪构成，在玻璃基板或者光纤端面直接制作生物分子膜，无需镀膜，制作简单，成本低廉。本发明无需测试透过待测溶液的信号，因此可以测试吸收大，甚至是不透明的液体。同时因为相位信息从干涉信号中提取，不受光源的波动和探测器灵敏度的影响，且不受到待测溶液折射率的影响。The label-free optical biosensor based on the cascaded adjustable periodic filter and Michelson interferometer provided by the present invention is composed of an adjustable periodic filter and a Michelson interferometer, and a biomolecular film is directly fabricated on a glass substrate or an optical fiber end face , without coating, easy to manufacture and low in cost. The present invention does not need to test the signal passing through the solution to be tested, so it can test the liquid with large absorption or even opaque. At the same time, because the phase information is extracted from the interference signal, it is not affected by the fluctuation of the light source and the sensitivity of the detector, and is not affected by the refractive index of the solution to be measured.
发明内容Contents of the invention
针对现有技术的不足，本发明的目的是提供一种成本低廉、结构简单、灵敏度高的基于可调周期性滤波器和迈克尔逊干涉仪级联的无标记光学生物传感器的技术方案。本发明使用低成本的宽带光源和滤波器的组合作为输入光源，探测器作为输出光的接收器。当待测溶液中被测物质吸附在生物分子膜后，引起生物分子膜增厚，导致迈克尔逊干涉仪的测量臂光程增加。记录被测物质吸附前后，探测器接收到的干涉光功率随可调滤波器光谱扫描变化的信息，经过傅里叶变换后，可以得到相位变化信息，推算出生物分子膜的厚度变化，从而获取待测溶液中被测物质含量信息。Aiming at the deficiencies of the prior art, the purpose of the present invention is to provide a low-cost, simple-structure and high-sensitivity technical solution based on a label-free optical biosensor cascaded with an adjustable periodic filter and a Michelson interferometer. The invention uses a combination of a low-cost broadband light source and a filter as an input light source, and a detector as a receiver of output light. When the substance to be measured in the solution to be measured is adsorbed on the biomolecular film, the biomolecular film is thickened, resulting in an increase in the optical path length of the measuring arm of the Michelson interferometer. Record the change information of the interference light power received by the detector with the spectral scanning of the adjustable filter before and after the adsorption of the measured substance. After Fourier transform, the phase change information can be obtained, and the thickness change of the biomolecular film can be calculated, so as to obtain Content information of the substance to be tested in the solution to be tested.
本发明解决其技术问题采用的技术方案是：一种基于可调滤波器和迈克尔逊干涉仪级联的光学生物传感器，包括宽带光源、可调周期性滤波器、迈克尔逊干涉仪和探测器；宽带光源的光经过可调周期性滤波器后进入迈克尔逊干涉仪；所述迈克尔逊干涉仪的其中一个臂的反射端面修饰可以吸附待测溶液中被测物质的生物分子膜，且与待测溶液接触，作为测量臂；另外一个臂的反射端面不修饰生物分子膜，且与待测溶液接触，作为参考臂；迈克尔逊干涉仪的输出端与探测器相连接。The technical solution adopted by the present invention to solve the technical problem is: a kind of optical biosensor based on the cascade connection of adjustable filter and Michelson interferometer, including broadband light source, adjustable periodic filter, Michelson interferometer and detector; The light of the broadband light source enters the Michelson interferometer after passing through an adjustable periodic filter; the reflective end surface modification of one of the arms of the Michelson interferometer can absorb the biomolecular film of the substance to be measured in the solution to be measured, and is compatible with the The solution contact is used as a measuring arm; the reflection end of the other arm is not modified with a biomolecular film and is in contact with the solution to be measured, which is used as a reference arm; the output end of the Michelson interferometer is connected to the detector.
进一步地，所述生物分子膜材料的折射率与被修饰的反射端面材料的折射率相同。Further, the refractive index of the biomolecular film material is the same as that of the modified reflective end surface material.
进一步地，通过调整所述迈克尔逊干涉仪的两个臂长，使得迈克逊干涉仪输出谱线的自由光谱范围与可调周期性滤波器输出光谱的自由光谱范围相同；当待测溶液中被测物质吸附在生物分子膜后，引起生物分子膜增厚，导致迈克尔逊干涉仪的测量臂光程增加。Further, by adjusting the two arm lengths of the Michelson interferometer, the free spectral range of the Michelson interferometer output spectrum is the same as the free spectral range of the output spectrum of the adjustable periodic filter; when the solution to be measured is After the substance to be measured is adsorbed on the biomolecular film, the biomolecular film is thickened, resulting in an increase in the optical path length of the measuring arm of the Michelson interferometer.
进一步地，所述可调周期性滤波器可以是法布里-珀罗谐振腔；或者是马赫-曾德干涉仪；或者是环形谐振腔；或者微盘谐振腔；所述可调周期性滤波器输出谱线谐振峰的位置可以通过改变滤波器的谐振腔长度进行移动。Further, the tunable periodic filter may be a Fabry-Perot resonator; or a Mach-Zehnder interferometer; or a ring resonator; or a microdisk resonator; the tunable periodic filter The position of the resonant peak of the output spectrum can be moved by changing the resonant cavity length of the filter.
进一步地，所述迈克尔逊干涉仪可以是由基于空间光学的分立器件构成；或者是基于光纤的结构构成；或者是基于集成光波导器件构成。Further, the Michelson interferometer may be composed of a discrete device based on space optics; or a structure based on an optical fiber; or based on an integrated optical waveguide device.
本发明具有的有益效果是：本发明光学生物传感器可利用宽带光源，无需测试光谱信息，不需要光谱仪，可以减小传感器的体积。在玻璃片、波导或者光纤端面修饰生物分子膜，无需在传感器的传感区域镀金属或者介质膜，大大降低了传感器的成本。测试信号是传感区域的反射光干涉信号，因此不用考虑光在待测液体中的吸收。通过测试迈克逊干涉仪传感臂的相位变化，获取生物分子膜厚度变化信息。因为相位信息从干涉信号中提取，不受光源的波动和探测器灵敏度的影响，且不受到待测溶液折射率的影响。The invention has the beneficial effects that: the optical biosensor of the invention can use a broadband light source, does not need to test spectral information, does not need a spectrometer, and can reduce the volume of the sensor. By modifying the biomolecular film on the glass plate, waveguide or optical fiber end face, there is no need to plate metal or dielectric film on the sensing area of the sensor, which greatly reduces the cost of the sensor. The test signal is the reflected light interference signal of the sensing area, so the absorption of light in the liquid to be tested is not considered. By testing the phase change of the sensing arm of the Michelson interferometer, the change information of the biomolecular film thickness is obtained. Because the phase information is extracted from the interference signal, it is not affected by the fluctuation of the light source and the sensitivity of the detector, and is not affected by the refractive index of the solution to be measured.
附图说明Description of drawings
图1为本发明中基于可调滤波器和迈克尔逊干涉仪级联的光学生物传感器的结构示意图；Fig. 1 is the structural representation of the optical biosensor based on tunable filter and Michelson interferometer cascaded among the present invention;
图2为可调滤波器在初始位置δL＝0以及δL＝534.5nm的输出谱线图；Fig. 2 is the output spectrum diagram of the adjustable filter at the initial position δL=0 and δL=534.5nm;
图3为迈克尔逊干涉仪在吸附待测物质前δh＝0以及吸附待测物质后δh＝267.2nm的输出谱线图；Fig. 3 is the output spectrum diagram of δh=0 and δh=267.2nm after absorbing the substance to be measured by Michelson interferometer;
图4为在可调周期性滤波器谐振腔长度为初始位置δL＝0时，待测物质吸附前后可调滤波器和迈克尔逊干涉仪级联后输出谱线图；Fig. 4 is when the resonant cavity length of the adjustable periodic filter is the initial position δL=0, the output spectrum diagram of the adjustable filter and the Michelson interferometer cascaded before and after the substance to be measured is adsorbed;
图5为待测物质吸附前后探测器接收到的光功率随可调滤波器谐振腔长度变化曲线。Fig. 5 is a graph showing the variation of optical power received by the detector with the length of the resonant cavity of the adjustable filter before and after the substance to be measured is adsorbed.
图6(a)为探测器接收到的干涉光信号傅里叶变换的频谱图。Fig. 6(a) is a spectrum diagram of the Fourier transform of the interference optical signal received by the detector.
图6(b)为探测器接收到的干涉光信号傅里叶变换的相位图。Fig. 6(b) is the phase diagram of the Fourier transform of the interference light signal received by the detector.
图7为傅里叶变换得到的相位信息随生物分子膜厚度变化图。Fig. 7 is a graph showing the change of phase information obtained by Fourier transform with the thickness of the biomolecular film.
具体实施方式detailed description
下面结合附图和实施例对本发明作进一步的说明。The present invention will be further described below in conjunction with the accompanying drawings and embodiments.
图1是本发明的第一个实施方式示意图。它包括：一个宽带光源(1)、一个可调周期性滤波器(2)、一个迈克尔逊干涉仪(3)、一个探测器(5)。所述迈克尔逊干涉仪(3)的其中一个臂(31)的反射端面(32)修饰了可以吸附待测溶液(4)中被测物质的生物分子膜(6)，且与待测溶液(4)接触，作为测量臂(31)；另外一个臂(33)的反射端面(34)不修饰生物分子膜，且与待测溶液(4)接触，作为参考臂；迈克尔逊干涉仪的输出端(35)与探测器(5)相连接。Fig. 1 is a schematic diagram of the first embodiment of the present invention. It includes: a broadband light source (1), an adjustable periodic filter (2), a Michelson interferometer (3), and a detector (5). The reflective end face (32) of one of the arms (31) of the Michelson interferometer (3) has modified the biomolecular film (6) that can adsorb the measured substance in the tested solution (4), and is compatible with the tested solution ( 4) contact, as a measuring arm (31); the reflective end face (34) of the other arm (33) does not modify the biomolecular film, and is in contact with the solution to be measured (4), as a reference arm; the output end of the Michelson interferometer (35) is connected with detector (5).
假设宽带光源(1)在1.53μm-1.57μm光谱是平坦的，每个波长对应的光场强度均为单位1。设可调周期性滤波器(2)由集成光波导环形谐振腔构成，输入和输出直波导与环形谐振腔的交叉耦合系数均为0.5，谐振腔长度为L＝400μm，谐振腔的有效折射率n1为1.45。在谐振腔在初始位置δL＝0以及δL＝534.5nm时可调周期性滤波器(2)的透射率曲线T1如图2所示。从图2中可见，因为谐振腔比较长，所以当谐振腔长度发生变化时，谐振腔的自由光谱范围没有发生明显变化，但是谐振波长位置发生了显著移动。Assuming that the spectrum of the broadband light source (1) is flat at 1.53 μm-1.57 μm, the light field intensity corresponding to each wavelength is unit 1. Assuming that the adjustable periodic filter (2) is composed of an integrated optical waveguide ring resonator, the cross-coupling coefficients of the input and output straight waveguides and the ring resonator are both 0.5, the length of the resonator is L=400 μm, and the effective refractive index of the resonator is n1 is 1.45. The transmittance curve T1 of the adjustable periodic filter ( 2 ) is shown in FIG. 2 when the resonant cavity is at the initial position δL=0 and δL=534.5nm. It can be seen from Figure 2 that because the resonant cavity is relatively long, when the length of the resonant cavity changes, the free spectral range of the resonant cavity does not change significantly, but the position of the resonant wavelength moves significantly.
本实例中迈克尔逊干涉仪是由2×2的光纤耦合器构成(分光比为50％：50％)，可调周期性滤波器(2)与光纤耦合器的一个输入端口相连接，迈克尔逊干涉仪的两个臂(31)和(33)分别为光纤耦合器的两个输出光纤，所以反射端面(32)和(34)是光纤端面，光纤耦合器的另外一个输入端与探测器(5)相连接。假设光纤等效折射率以及生物分子膜的折射率n1均为1.45；待测溶液的折射率n2为1.33。In this example, the Michelson interferometer is composed of a 2×2 fiber coupler (the splitting ratio is 50%: 50%), and the adjustable periodic filter (2) is connected with an input port of the fiber coupler, and the Michelson The two arms (31) and (33) of the interferometer are respectively two output fibers of the fiber coupler, so the reflective end faces (32) and (34) are fiber end faces, and the other input end of the fiber coupler is connected to the detector ( 5) Connected. It is assumed that the equivalent refractive index of the optical fiber and the refractive index n 1 of the biomolecular film are both 1.45; the refractive index n 2 of the solution to be measured is 1.33.
在光纤端面或生物分子膜表面与待测溶液的反射率为RThe reflectance between the optical fiber end face or the surface of the biomolecular film and the solution to be measured is R
因为迈克尔逊干涉仪的两个臂的端面反射率相同，所以迈克尔逊干涉仪的输出谱线为T2 Because the end reflectivity of the two arms of the Michelson interferometer is the same, the output line of the Michelson interferometer is T 2
其中h为迈克尔逊干涉仪(3)两臂的光程差，为使迈克逊干涉仪(3)输出谱线的自由光谱范围与可调周期性滤波器(2)输出光谱的自由光谱范围相同h＝800μm，δh为生物分子膜吸附被测物质后引起的生物分子膜厚度变化，λ为入射光波长。图3给出了迈克尔逊干涉仪在吸附前δh＝0以及吸附后δh＝267.2nm的输出谱线T2。因为两个臂的反射率相同，所以透射率的最小值可以等于0。从图3中可见，因为迈克尔逊干涉仪两臂的光程差比较长，所以当待测物质发生吸附时，迈克尔逊干涉仪的自由光谱范围没有发生明显变化，但是干涉极值波长位置发生了显著移动。in h is the optical path difference of the two arms of the Michelson interferometer (3), in order to make the free spectral range of the Michelson interferometer (3) output spectral line the same as the free spectral range of the adjustable periodic filter (2) output spectrum h =800μm, δh is the thickness change of the biomolecular film caused by the biomolecular film adsorbing the measured substance, and λ is the wavelength of the incident light. Fig. 3 shows the output spectrum T 2 of the Michelson interferometer before adsorption at δh=0 and after adsorption at δh=267.2nm. Since the reflectivity of both arms is the same, the minimum value of the transmittance can be equal to 0. It can be seen from Figure 3 that because the optical path difference between the two arms of the Michelson interferometer is relatively long, when the substance to be measured is adsorbed, the free spectral range of the Michelson interferometer does not change significantly, but the wavelength position of the interference extremum changes. Move significantly.
图4为在可调滤波器谐振腔长度为初始位置δL＝0时，待测物质被吸附前后可调周期性滤波器和迈克尔逊干涉仪级联后输出谱线图。在待测物质被吸附前，可调周期性滤波器和迈克尔逊干涉仪的极大值和极小值位置均重合，此时级联后透射率最大；在待测物质被吸附后δh＝267.2nm，可调周期性滤波器的极大值和迈克尔逊干涉仪的极小值位置重合，可调周期性滤波器的极小值和迈克尔逊干涉仪的极大值位置重合，此时级联后透射率最小。Fig. 4 is an output spectrum diagram of the adjustable periodic filter and the Michelson interferometer cascaded before and after the substance to be measured is adsorbed when the length of the resonant cavity of the adjustable filter is at the initial position δL=0. Before the substance to be measured is adsorbed, the positions of the maximum and minimum values of the adjustable periodic filter and the Michelson interferometer coincide, and the transmittance is the largest after cascading; after the substance to be measured is adsorbed, δh=267.2 nm, the maximum value of the adjustable periodic filter coincides with the minimum value of the Michelson interferometer, and the minimum value of the adjustable periodic filter coincides with the maximum value of the Michelson interferometer. At this time, the cascade The rear transmittance is minimal.
当可调周期性滤波器的谐振腔变化范围从-6μm到6μm时(假设扫描一个周期用12ms)，在待测物质被吸附前后，探测接收到的光功率变化如图5所示。可见当待测物质被吸附时，光功率变化曲线发生了移动，将光功率变化曲线进行傅里叶变换，计算主频的相位变化信息，就可测出吸附待测物质后生物分子膜厚度变化。When the resonant cavity of the adjustable periodic filter changes from -6 μm to 6 μm (assuming that a scan cycle takes 12 ms), before and after the substance to be measured is adsorbed, the change of the detected optical power is shown in Figure 5. It can be seen that when the substance to be tested is adsorbed, the optical power change curve moves, and the optical power change curve is Fourier transformed, and the phase change information of the main frequency is calculated to measure the thickness change of the biomolecular film after the substance to be tested is adsorbed. .
图6是待测物质被吸附前后，探测器接收到的光功率变化曲线的傅里叶变换信息。当待测物质被吸附前后，其主频率的位置不变(均在916.6Hz)，但是相位随着生物分子膜吸附待测物质的厚度变化而变化。如图7所示，相位从π到π/2，再到0，分别对应吸附厚度从初始状态0nm，到133.6nm，到267.2nm。生物分子膜的厚度每增加1nm，相位减少0.0118rad。当入射光源的功率发生变化，或者被测样品折射率发生变化时，影响的仅仅是探测器接收到的功率的波动，但是其相位并不受到影响，因此仍然可以测出被测物质含量。Fig. 6 is the Fourier transform information of the optical power change curve received by the detector before and after the substance to be measured is adsorbed. Before and after the substance to be tested is adsorbed, the position of its main frequency remains unchanged (both at 916.6 Hz), but the phase changes with the thickness of the biomolecular membrane adsorbing the substance to be tested. As shown in Figure 7, the phase changes from π to π/2, and then to 0, corresponding to the adsorption thickness from the initial state of 0 nm, to 133.6 nm, to 267.2 nm, respectively. For every 1nm increase in the thickness of the biomolecular film, the phase decreases by 0.0118rad. When the power of the incident light source changes, or the refractive index of the measured sample changes, only the fluctuation of the power received by the detector is affected, but its phase is not affected, so the content of the measured substance can still be measured.
上述实施例用来解释说明本发明，而不是对本发明进行限制。在本发明的精神和权利要求的保护范围内，对本发明作出的任何修改和改变，都落入本发明的保护范围。The above-mentioned embodiments are used to illustrate the present invention, but not to limit the present invention. Within the spirit of the present invention and the protection scope of the claims, any modification and change made to the present invention will fall into the protection scope of the present invention.
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