光纤端的等离激元探测技术 
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1 引言
在单模光纤的端平面集成表面等离激元谐振(SPR)微纳器件,是融合微纳光学与光纤通信技术的一种独特途径。在对生化物质和环境场的探测中,这类技术已经清晰地展示出应用价值。与基于自由空间光束耦合到表面等离极化激元(SPP)的现有商业设备相比,光纤导波耦合的方式具有体积紧凑、光路灵活的显著优势。同时,细小的光纤端可以插进微量样品通过dip-and-read的方式直接读取信号,也能够伸入狭小的空间进行内窥探测,减轻内窥操作对检测对象造成的伤害。近10年来,针对这类器件,人们不断地发展与提升理论模型及制备工艺,并演示了对生物分子与声波的探测功能。在2018年英国物理学会发布的报告《The health of photonics》中,针对老龄化社会列出6项光学技术解决方案,其中就包括“在专用光纤上安装微小传感器,实现即时、高灵敏度的化学与生物分析。”[1] 另一方面,锥形光纤探针,因其较低的背景散射和便捷的制备工艺,也是一种集成等离激元纳米器件的良好平台。其中,通过扫描探针显微(SPM)技术操纵黏附在锥形光纤顶端的等离激元纳米颗粒天线,是实现等离激元天线精密动态调控的重要手段。本文拟结合作者的研究心得,对这两方面的研究现状、发展路径和应用前景进行介绍与讨论。
2 单模光纤端面集成表面等离激元谐振探测技术
在单模光纤上集成较高灵敏度的SPR传感器,具有低损伤侵入式测量、通过柔软细小的光纤传导信号、结合成熟的光纤通信技术、紧凑稳定的光学系统等诸多技术能力和优势。而其所采用的dip-and-read简单操作方式大大减少了操作时间和复杂度,这对SPR技术往急诊、床边检验、现场检验等应用领域发展具有重要的价值。Lab-on-fiber已经发展成微纳光学领域的一个重要研究方向[2-8]。以下先从结构设计与制备工艺两方面介绍若干代表性成果,然后介绍生物分子与声波探测应用的发展情况,最后讨论未来可能的发展方向及需要注意的问题。
2.1 表面等离激元谐振的结构设计与理论模型
目前其他研究团队报道的单模光纤端面SPR结构基本都延续了自由空间平面光波与芯片上的SPR结构通过光栅耦合的思路,采用单一的周期性结构,或者较少考虑各个SPR单元的相互耦合而注重独立单元与光波的耦合性能,例如纳米颗粒阵列、纳米孔阵列、光栅和超构表面等[2,5-6,9-24]。
为避免单模光纤与端面SPR结构耦合的困难,有较多的研究工作转而采用多模光纤。
图 1. 光纤与SPR集成器件示例。(a)单模光纤端面单一周期性结构[13];(b)多模光纤端面器件[14];(c)光纤侧壁器件[29];(d)单模光纤端面SPR微腔[25]
Fig. 1. Examples of optical-fiber and integrated SPR devices. (a) Uniform periodic structure on single-mode optical fiber's end-facet[13]; (b) device on multi-mode optical fiber's end-facet[14]; (c) device on optical fiber's sidewall[29]; (d) SPR microcavity on single-mode optical fiber's end-facet[25]
另有大量的研究工作将SPR结构放在光纤侧壁,同样可以避免单模光纤与端面SPR结构耦合的困难[2,4-5,28-43]。
2016年以来,为提高单模光纤导波和端面SPR结构耦合的
图 2. 单模光纤端面SPR微腔结构。(a)(b) SPP带边态结构示意图及靠近结构中央处的扫描电子显微镜照片[25];(c)(d) SPP禁带内缺陷态结构示意图及靠近结构中央处的扫描电子显微镜照片[25]
Fig. 2. SPR microcavities on single-mode optical fiber end-facets. (a)(b) Schematic of a structure for achieving an intraband SPP cavity mode and its SEM image near the center of the structure[25]; (c)(d) schematic of a structure for achieving a defect cavity mode in the SPP bandgap and its SEM image near the center of the structure[25]
图 3. SPR微腔谐振态的理论与实验结果。(a) 645 nm周期无穷大纳米槽阵列的SPP能带计算图[44];(b) SPP禁带(点划线)内缺陷态(箭头)及其随缺陷宽度s变化的计算图[44], s定义见图2(c);(c)单模光纤端面SPP带边态结构的反射谱[25]
Fig. 3. Theoretical and experimental results for SPR microcavity resonant state. (a) SPP band diagram for infinitely wide nanoslit array with period of 645 nm[44]; (b) defect modes (arrows) in the SPP bandgap (dash-dot lines), and their dependence on the defect's width s, refer to Fig. 2(c) for the definition of s[44]; (c) reflection spectra of an intraband SPP cavity mode structure on single-mode optical fiber's end-facet[25]
2.2 制备工艺
在微细的光纤端面进行高质量和精确对准的纳米结构加工对制备工艺提出了挑战。产业化的制备工艺要求高效率、高良品率和高可重复性。人们尝试了在光纤端面直接用电子束光刻[13-14]、离子束刻蚀[9,16-18]等方法加工SPR结构,及各种把SPR结构转移到光纤端面的方法,包括nanoskiving[12]、decal transfer[10]、template stripping[25,47-49]等。
2016年,本课题组报道了将平面玻璃衬底上的SPR微腔结构精确转移到单模光纤端面的制备工艺,如
图 4. 光纤端面SPR结构的3种转移加工技术。(a) Nanoskiving[12];(b) decal transfer[10];(c) glue-and-strip[25]
Fig. 4. Three transfer techniques for fabricating SPR structures on optical fiber end-facets. (a) Nanoskiving[12]; (b) decal transfer[10]; (c) glue-and-strip[25]
2.3 生物分子相互作用分析应用
目前,SPR传感器的最主要应用是检测和分析生物大分子之间的结合和解离实时动态过程。当折射率LOD达到一定水平后,实际的分子探测下限和良好的动力学表征性能并不单纯取决于甚至不主要取决于折射率灵敏度,必须综合测量噪声、环境控制、流体力学、化学表面、配体固定等因素进行全面的设计才能得到好的实验结果。对相关内容我们将另行撰文介绍。
在前期工作中,本课题组以多聚赖氨酸为骨架的聚乙二醇(PEG)为亲水表面,以生物素(biotin)为配体进行了一些简单的验证性实验。
图 5. 基于单模光纤端面SPR微腔传感器的生物分子相互作用分析实验[7]。其中除了第一步基线测试外,每步都是先浸入分子溶液,再浸入缓冲液,两个区间以曲线上的一个小尖为分界
Fig. 5. Biomolecule interaction analysis experiment based on SPR microcavity sensor on single-mode optical fiber's end-facet[7]. Except for the first step which is baseline, each step comprises of two parts which are immersing the sensor in the molecule solution and then in the buffer solution. There is a little spike on the testing curve between the two parts
2.4 超声探测应用
基于声压改变折射率的声光效应,SPR也被用于超声探测[50-51]。虽然目前其灵敏度远低于基于高
针对压电水听器灵敏度和带宽的矛盾,光学谐振微腔提供了一个有前景的超声探测方案,在过去约20年间得到了较多的研究[52-65]。其基本的声光转换原理是,将固定波长激光耦合到微腔,以声压改变微腔的等效折射率或几何尺寸,引起微腔谐振波长的移动,从而导致激光反射或透射功率随声压的变化。基于高
图 6. 典型微纳声光器件示意图(它们的光学导波功能与声光转换功能由同一种材料完成)。(a)微环[54];(b)法布里-珀罗腔[62]
Fig. 6. Schematics of typical acousto-optic micro devices (in both devices, optical waveguiding and acousto-optic transduction are performed by the same material). (a) Micro-ring[54]; (b) Fabry-Pérot cavity[62]
图 7. 以单模光纤端面SPR微腔对超声进行探测[51]。(a)实验系统示意图;(b)SPR反射谱;(c)对10 MHz中心频率超声脉冲的反射光功率响应
Fig. 7. Ultrasound detection with an SPR microcavity on a single-mode optical fiber end-facet. (a) Schematic of the experimental system; (b) SPR reflection spectrum; (c) undulation of laser power reflection in response to a series of ultrasound pulse with a central frequency of 10 MHz
尽管微环等水听器达到了很高的声探测性能,但对需要极小器件尺寸的内窥场景而言,将微腔直接集成在单模光纤的端面是更加具有高价值应用前景的方式。英国伦敦大学学院在此方面进行了多年的研究并不断取得突破[61-64]。2009年,他们报道了单模光纤端面的平面镜法布里-珀罗(F-P)腔,其多聚物薄膜腔体的长度随声压变化,得到NEP为5 kPa(20 MHz内),带宽为50 MHz[61]。这种器件已经商业化,是Precision Acoustics公司的UMS3超声探测系统的核心技术。之后,通过利用表面张力来制备球面镜F-P腔,如
基于紫外固化胶折射率随声压的变化,单模光纤端面SPR微腔也具有超声探测能力。我们将光纤导波耦合到沿金薄膜朝向固化胶一面的SPR谐振模式,工作波长为1550 nm附近[51]。如
2.5 未来发展方向和关键技术突破点
对SPR技术的生物分子检测应用而言,互作动力学分析是当前最大的市场,已经广泛应用于生命科学研究和药物研发,全球市场规模10亿美元左右[66-67]。在此应用领域,光纤端面SPR传感技术因其方便、快速和小型化的特点,具有取代当前基于自由空间光学和微流控进样的大型、复杂SPR仪器的巨大潜力。进一步,如果能够实现对粗样品中生物标志物含量的快速、精准定量测量,显著提高床旁免疫诊断和农产品检验的准确性和速度,这将把SPR技术应用带入前所未有的广阔天地。基于dip-and-read方式对微量粗样品进行方便、快速的检测,将成为光纤端传感器在未来应用上的一大优势。
需要注意的是,当折射率LOD达到10-6 RIU或更低水平后,在传感器研究层面,如何去除环境和杂质干扰成为比灵敏度更关键的问题,直接限制着真实的探测限。以对心肌肌钙蛋白的高敏检测为例,欧洲心脏病学学会指南要求对健康人群第99百分位的定量误差低于10%[68],经计算可知这相当于随机信号漂移应小于0.01 pg·(mm-2·min-1)[假设满吸附信号为1000 pg·mm-2,动力学结合常数为106 M-1·s-1 (1 M=1 mol/L),标志物浓度0.02 μg/L,标志物分子量约为20 kDa],这个要求已经高于Biacore 8K产品手册给出的扣减空白参照后的漂移值。未关注环境和杂质干扰的极低含量探测是没有意义的,即便在实验中看起来获得了可重复的结果也需要审慎对待,相关内容我们会另行撰文介绍。
对声信号检测应用而言,用较低
因此,我们认为,通过增加材料的声光响应而非增加微腔的光学
3 锥形光纤端局域表面等离子体谐振天线探测技术
此章节中,我们将讨论以贵金属纳米颗粒作为等离激元天线传感器,测量衬底表面性质的工作。具体地,我们将首先介绍由一对金属纳米颗粒或单颗粒及其镜像组成的dimer等离激元天线,然后介绍对等离激元天线进行动态调控的意义和此领域内非光纤集成技术的研究进展,最后讨论将等离激元天线和扫描显微技术及锥形光纤进行集成以实现动态调控的工作。
3.1 贵金属纳米颗粒-镜像等离激元天线
类似于电感电容回路接收特定波长的无线电波,等离激元天线能够形成局域表面等离子体谐振(LSPR),在深亚波长尺度对光波进行有效的散射、吸收和聚焦[72-73]。而当一对贵金属纳米颗粒互相靠近,组成一个dimer LSPR天线时,在夹在两个颗粒中间的间隙处可形成纳米至亚纳米尺度的hotspot,在谐振波长处hotspot内的电场能量密度可比入射光提高几百至上万倍[74-79]。如此小尺寸和高态密度的hotspot,提供了以纳米尺度分辨率探测极大增强的光与物质相互作用的手段,探测对象包括分子吸附或变形、分子振动谱、光电转换、非线性混频、荧光和量子隧穿等[75-99]。
精细且可控地调节dimer LSPR天线的间隙距离对控制天线谐振谱及hotspot与物质的作用强度非常重要。目前最可控、可重复的方法之一是将贵金属纳米颗粒置于一个镜面上,使颗粒与其镜像形成dimer LSPR天线,而间隙宽度由颗粒与镜面之间的物质厚度决定,比如若干单分子层或若干层二维材料[76,95,100-103]。近年来,科研工作者们在这种体系中观察到了大Purcell因子、奇异电致发光、单分子电致单光子源、荧光Rabi劈裂、单分子级别化学过程、强非线性光学、非局域和量子隧穿效应、皮米级hotspot的产生和湮灭,及可能来自于声子受激辐射的非线性拉曼散射等众多新奇现象[79,86,94-95,104-112]。
通过聚焦空间径向偏振的激光来激发这种贵金属纳米颗粒-镜像等离激元天线的纵向偏振LSPR,在60 nm直径金球-单层分子-原子级平滑金面的体系中,在约3 nm直径的hotspot里得到超过109的拉曼电磁增强因子,同时各个hotspot的增强因子高度可重复,如
图 8. 聚焦空间径向偏振的激光激发60 nm直径金球-单层分子-原子级平滑金面体系[76]。(a)示意图,画出了金球的镜像;(b)在纵向LSPR谐振条件下,hotspot纵向电场能量密度分布的仿真结果,颜色表示相对于入射光的增强倍数;(c)吸附了单层4-nitrobenzenethiol分子的20个不同的金球在300 nW入射激光功率下的拉曼光谱,展示了极大增强倍数的可重复性
Fig. 8. Focusing radially polarized laser beam to excite structure which contains a 60-nm gold nanosphere, a monolayer of molecules, and an atomically flat gold surface[76]. (a) Schematic, with the gold nanosphere's mirror image; (b) simulation result for the vertical electric field component's intensity distribution in the hotspot under vertical LSPR resonance, with color scale indicating enhancement compared to the incident light; (c) experimental Raman spectra of 20 different gold nanospheres, each c
3.2 动态调控的等离激元天线
如果能够使贵金属纳米颗粒的位置在镜面上移动甚至得到精确的动态控制,将带来关于LSPR天线更丰富的科学研究内容和更广泛的技术应用范围。例如,LSPR谐振波长随间隙高度的灵敏变化被用来测量间隙内分子的大小变化[115-117],
进一步,如果能够将贵金属纳米颗粒集成在SPM的针尖,对间隙尺寸实现纳米和亚纳米尺度的调控,同时对样品实现扫描成像,将大大提高LSPR天线探测技术的能力。在应用于增强拉曼散射时,这相当于表面增强和针尖增强两种技术的结合,既具有LSPR带来的更强的拉曼信号,又具有SPM技术的高空间分辨率。在此方向,中国科学技术大学实现了0.5 nm空间分辨率的单分子拉曼成像,单分子与LSPR的耦合调控,及若干分子间的电偶极矩耦合调控与测量,如
图 9. 动态调控的等离激元天线。(a)银纳米线-镜面结构的LSPR波长随间隙内分子的热胀冷缩而移动[117];(b)基于DNA折纸术控制单个荧光分子在hotspot中的位置[119];(c)用气凝胶上的金纳米球对聚焦光斑的空间偏振态进行扫描成像,右下子图显示成像结果[120];(d)STM结合LSPR效应对单个和若干个分子进行调控和测量(上两行是单个分子与等离激元的Fano耦合,底行是对两个分子电偶极矩耦合的扫描成像)[123-124]
Fig. 9. Dynamically tuned plasmonic antennas. (a) The LSPR of a silver nanowire-mirror structure is tuned by thermal expansion of molecules in the gap[117]; (b) the relative position of a single fluorescence molecule in a plasmonic hotspot is controlled by DNA origami[119]; (c) the spatial polarization state of a laser focal spot is imaged by scanning a gold nanosphere on aerogel, with an imaging result in the inset; (d) tuning and measurement of single and few molecules using an LSPR probe on STM (top
迄今,人们已发展出多种集成在SPM针尖的等离激元结构[125-141]。
图 10. 集成在SPM针尖的等离激元结构。(a)在AFM针尖用聚焦离子束刻蚀制作的LSPR天线[128];(b)一对AFM针尖上的金球组成dimer LSPR天线,通过导电AFM控制和测量天线间隙[96];(c)AFM探针上雕刻的金属光栅将入射光波耦合到SPP,并将SPP聚焦到针尖[133];(d)雕刻了螺旋形金光栅的AFM探针具有手性光力作用[139];(e)基于锥形光纤与银纳米线高效率耦合的近场光学显微镜探针[141]
Fig. 10. Plasmonic structures integrated on SPM probes. (a) An LSPR antenna on an AFM probe apex has been fabricated using focused ion beam milling[128]; (b) a pair of gold spheres on AFM probe apexes comprise a dimer LSPR antenna, with its gap size controlled and measured by conductive AFM[96]; (c) a metallic grating carved on an AFM probe couples incident light waves to SPPs, and focuses SPPs to the probe apex[133]; (d) a helical gold grating carved on an AFM probe is used to detect enantioselective op
3.3 锥形光纤探针尖端的等离激元天线
AFM探针上雕刻SPR结构的方法,具有一定的工艺难度,而且这种SPR结构的表面粗糙度不如化学合成的金属纳米颗粒,有可能影响hotspot的分布和可控性。因此,把化学合成的等离激元天线黏附到探针尖端是一种有益的集成SPM技术与LSPR天线的方式。其中一个典型方法是先用氢氟酸腐蚀或其他手段形成锥形光纤探针,在光纤探针上附着黏性材料,然后以光纤探针对平面衬底上的金属纳米颗粒作AFM扫描并将金属纳米颗粒粘到光纤探针的尖端,最终形成以金属纳米颗粒为探针的AFM扫描显微技术[143-144]。其中,光纤可以固定在音叉上,通过将音叉的振动转换为电信号来实现原子力反馈[145]。
在
图 11. 基于锥形光纤尖端金纳米颗粒探针的AFM技术。(a)金纳米颗粒探针接近另一个金纳米颗粒时,四波混频信号的急遽上升[146];(b)金纳米颗粒探针接近单个分子时,分子荧光从增强到淬灭的过程[147]
Fig. 11. AFM technology based on gold nanoparticle probes on tapered optical fibers' apexes. (a) As a gold nanoparticle probe approaches another gold nanoparticle, the four-wave-mixing signal increases significantly[146]; (b) as a gold nanoparticle probe approaches a single molecule, the molecular fluorescence experiences the process from enhancement to quenching[147]
虽然锥形光纤的折射率较小,但其线性散射相对于金属纳米颗粒的LSPR散射来说仍然是一个较大的背景,因此在前期各篇报道里也未对锥形光纤上金属纳米颗粒探针的LSPR散射光谱进行有效的测量[148-149]。最近,本课题组通过空间和偏振滤波的方法提取纵向LSPR模式的散射,并通过数学拟合扣除锥形光纤的背景散射,实现了对锥形光纤尖端100 nm金球LSPR散射谱的测量,并根据LSPR谐振波长的移动得到了对玻璃衬底上100 nm金球的扫描图像,如
图 12. 以锥形光纤尖端100 nm金球的LSPR散射波长为待测变量,对玻璃衬底上的100 nm金球进行AFM扫描成像[150]。(a)金纳米球探针接近另一个金纳米球时,LSPR散射谱里纵向dimer模式的出现;(b)以金纳米球探针作AFM线扫描的形貌结果(上)和LSPR结果(下)
Fig. 12. Scanning LSPR microscopy of a 100 nm gold nanosphere on a glass substrate, using a 100 nm gold nanosphere on a tapered optical fiber's apex as the AFM probe, and its LSPR scattering spectrum as the imaged quantity[150]. (a) Schematic showing that as the gold nanosphere probe approaches a gold nanosphere target, a vertical dimer mode appears in the LSPR scattering spectrum; (b) line scanning result for morphology (top) and LSPR (bottom)
3.4 展望
在AFM技术的基础上,迄今人们已经发展出了对多种物理量的高空间分辨率测量方法,包括几何形貌、光场、磁场、静电场、力、温度、弹性模量、电容、压电响应和拉曼散射等。每一种新功能的实现都依赖于新型探针的发明。LSPR天线自身具有对表面环境折射率、分子吸附、拉曼散射、非线性光学、荧光和量子效应等多个参数或过程的高灵敏度响应。而锥形光纤端的LSPR天线,因其简便的制备方法、潜在的天线种类多样性和较低的背景散射,在SPM探测技术的未来发展中将有所作为。
4 结束语
结合光纤光学或SPM的光纤端等离激元探测技术,在科研、产业和经济等领域展示出引人入胜的前景。本文对其发展历程进行梳理,并在此基础上,结合作者的科研实践,对未来发展的重点、关键和潜在价值进行了讨论。
首先,对于光纤集成的SPR生物分子传感器,未来工作的中心之一是深入挖掘其方便、快速的核心价值,从而取代当前体积庞大、操作复杂的高端实验室仪器。在这方面,单模光纤端面SPR微腔以简便的dip-and-read测试方式、能够检验微量样本的探针、较高的灵敏度和稳定性、高效率的转移制备工艺等特点,对应用需求形成良好的响应。在此基础上,未来技术发展的关键是如何相对提高对目标分子的感应,及相对降低环境和杂质的干扰,从而解决复杂样品中低含量分子检测的难题。
其次,对于光纤集成SPR结构的声信号探测应用,如何基于SPP表面波的空间开放性,大大提升SPR对声信号的灵敏度,有可能是实现小型化、高灵敏度光纤水听器阵列的重要突破口。
最后,以锥形光纤尖端的LSPR天线为探针的SPM技术,能够以同一个LSPR天线或plasmonic hotspot对样品进行高精度调控和高空间分辨率测量,从而大大增加了对LSPR天线和其他对象之间相互作用的表征能力,同时也消除了基于固定天线的实验中不同天线之间hotspot差异对测量结果的影响,从而显著降低实验结果分析的复杂度。迄今,人们已经通过这类或类似技术实现了对分子发光、分子振动、非线性光学、量子隧穿、手性光力和线性LSPR散射等多种现象的测量。未来,结合LSPR探针结构的多样化设计,将能够实现更多的测量应用。
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Article Outline
杨天, 陈成, 王晓丹, 周鑫, 雷泽雨. 光纤端的等离激元探测技术[J]. 激光与光电子学进展, 2019, 56(20): 202404. Tian Yang, Cheng Chen, Xiaodan Wang, Xin Zhou, Zeyu Lei. Plasmonic Sensing on Fiber Tip[J]. Laser & Optoelectronics Progress, 2019, 56(20): 202404.
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