纳米分辨相干反斯托克斯拉曼散射显微成像

采用附加探测光声子耗尽法来实现超衍射极限相干反斯托克斯拉曼散射显微成像. 此方法引入一束环形分布的附加探测光来消耗点扩展函数周边的相干声子, 实现点扩展函数的改造, 从而达到超越衍射极限的空间分辨率. 为了获得更高的空间分辨率和更佳的相位匹配条件, 通常需采用高数值孔径物镜对抽运光、斯托克斯光和探测光进行聚焦, 此时标量衍射理论不再成立. 基于矢量衍射理论, 分析了线偏振光、圆偏振光先后经过螺旋相位片和高数值孔径物镜后的光强分布, 结果表明: 圆偏振光在高数值孔径物镜后焦平面的光强分布呈中心对称状, 较线偏振环形光更适合作为附加探测光. 此外, 采用全量子理论分析了附加探测光声子耗尽法. 结果表明: 当附加探测光与探测光强度比为80时, 成像系统的横向空间分辨率可以达到45 nm; 继续提高附加探测光强度, 空间分辨将进一步提高.     

共焦显微系统中光学超分辨光瞳滤波器的设计

利用矢量衍射理论设计了适用于共焦显微等大数值孔径系统的三区二元相位型超分辨光瞳滤波器.数值模拟了等效数值孔径为1.4的共焦显微系统焦点附近的光强分布,根据系统所需要的斯特雷尔比和轴向分辨率,采用优化搜索算法求解了能够实现光学超分辨的三区二元相位型光瞳滤波器结构.结果表明,利用所设计的三区二元相位型光瞳滤波器可以在较低旁...     

基于点扫描的超分辨显微成像进展

光学显微镜一直推动着现代科学技术的发展.随着科学的进步,对显微成像分辨率的要求在生物、材料等领域日渐凸显,而常规宽场显微成像一直面临着成像分辨率衍射受限的问题.1968年出现的共聚焦显微镜作为点扫描显微镜的开端第一次实现了远场下成像分辨率的突破,它具有层切性好、信噪比高等优点.在1994年出现的受激辐射荧光损耗显微镜将显微成像能力突破到2.8 nm左右,并成为目前效果最佳、应用较广泛的超分辨显微技术.荧光差分显微和饱和荧光吸收竞争等点扫描技术具有无荧光染剂限制、饱和光强低、光路简单等优势,并且能取得1/6波长的分辨能力,进而在超分辨显微领域仍有着发挥空间.Airyscan技术作为以上方法的补充可以弥补点扫描系统中由于探测小孔半径减小而带来的信号丢失,从而提高成像信噪比和分辨率,但阵列探测器成本较高.上述点扫描显微镜通过改变照明或者探测的方式实现了分辨率突破.本文详细讨论了点扫描超分辨方法的原理、成像效果及面临的瓶颈,并分析了点扫描超分辨显微镜在应用和技术上的趋势.     

随机光学重构显微成像技术及其应用

光学显微成像技术在生命科学、生物医学、临床医学诊断和材料科学等领域有着非常广泛的应用。但由于光学衍射极限的存在,传统光学显微镜无法观察到纳米尺度的物质及生命活动,极大地限制科学研究和医学的发展。近年来,随着突破光学衍射极限的超分辨成像技术的不断发展,显微成像分辨率得到不同程度的提高。目前在基于不同原理的各种超高分辨率显微镜中,随机光学重构显微镜(STORM)分辨率最高,可达几十纳米,真正实现了单分子水平检测。着重介绍了STORM超分辨显微成像技术的原理、实验方法及其应用。     

结构光照明超分辨光学显微成像技术与展望

结构光照明显微镜(Structured Illumination Microscopy,SIM)通过结构化照明在频率域以空间混频的方式将物体高频信息载入光学系统的探测通带内实现突破衍射极限的超分辨光学显微成像。SIM凭借其较低的激发光强、对荧光染料的非特异性需求以及快速的宽场成像优势已成为活细胞超分辨光学显微成像方面应用最多的技术。本文系统回顾了SIM的技术进展,对SIM的基本原理与实现方法进了详细的分析,重点介绍了本课题组研发的基于光谱分辨的单光子激发超分辨显微镜和结合自适应光学的双光子激发超分辨显微镜这两种最新的SIM技术,最后简要讨论了SIM技术在生物成像中的应用及未来发展方向。     

微球透镜超分辨显微成像与检测技术综述

受衍射极限的影响,传统光学显微镜的分辨率最高约为波长的一半,突破衍射极限,获得更高的成像分辨率是近年来显微成像领域的研究热点。相比于其他超分辨显微成像方式,基于微球透镜的超分辨显微成像方式具有简单直接、免标记等优点。主要介绍国内外研究团队将微球与传统的光学显微镜结合实现超分辨显微成像的研究进展,从微球透镜参数选择、成像方案、成像分辨率、成像视场及成像机理等多角度进行总结与比对;并结合课题组工作,介绍了将微球透镜与干涉显微技术相结合的三维超分辨检测技术,阐述了Linnik型与Mirau型两种检测光路原理,分析了三维超分辨检测的效果;展望了微球透镜超分辨显微技术在显微成像与显微干涉检测两个方面待解决的问题与发展方向。     

图像复原式整形环形光横向超分辨共焦显微测量新方法

提出一种新的图像复原式整形环形光横向超分辨共焦显微测量法. 该方法首先利用二元光学器件，将高斯照明光束整形为环形光束，用于初步改善共焦显微镜的横向分辨力，然后利用基于最大似然估计法(maximum likelihood estimate, MLE)的单幅图像超分辨复原技术，重建测量图像的高频信息，来进一步改善共焦显微镜的横向分辨力. 实验表明,当λ=632.8nm，N.A. =0.85时，该方法能使共焦显微镜获得优于0.1μm的横向分辨力. 利用该方法建立的横向超分辨共焦显微系统除了具有显著的超分辨效果外 关键词： 超分辨 超分辨复原 最大似然估计 共焦成像     

基于环形光瞳提高荧光辐射差分超分辨显微镜的成像分辨率

荧光辐射差分(FED)显微术利用实心光斑扫描的一幅共聚焦图像减去空心光斑扫描的负共聚焦图像,实现了超分辨成像。使用简单的环形光瞳滤波器提高了无变形FED的成像分辨率。在空心焦斑光路上使用合适的环形光瞳滤波器,压缩空心焦斑的尺寸;同时,在实心焦斑的光路上允许使用更高数值孔径的孔径光阑,获得更小的、与空心焦斑相匹配的实心光斑分布:两方面作用下,提高了FED的空间分辨率。     

基于近场光学的微球超分辨显微效应

在光学成像领域,由于受到衍射极限的限制,常规成像分辨率在200nm左右.科学的不断进步对更高分辨率有着迫切需求,如何突破这个极限来获得更高质量的高分辨率图像是热门研究领域.2011年提出了微球超显微技术:在原有的光学系统中,将直径几微米至几十微米的透明微球直接置于样品表面,就能够成倍提高传统光学显微镜的成像能力.微球超显微技术以其简单直接的特点,受到广泛关注.本文介绍了光学显微镜的研究背景以及国内外团队在微球超分辨显微技术方面的研究进展,包括通过在微球表面进行环刻同心环、中心遮挡和表面涂覆的方法来调节微球所产生的光子纳米喷射方面所开展的一系列研究,并进行了理论模拟和实验验证,进一步提升了微球的超分辨显微效应.最后,展望了今后微球超分辨显微技术的应用与发展方向.     

光学移频超分辨成像技术进展

光学显微镜具有无损、样品友好、速度快等优点,一直是人类探索微观世界的主要手段.但是,由于受到衍射极限限制,长期以来,光学成像系统的分辨率最高仅能达到可见光半波长量级,逐渐成为科学技术发展的桎梏.对于荧光标记样品,可以利用荧光超分辨光学显微成像技术打破光学衍射极限,填补电子显微镜(约为1 nm)和普通可见光学显微镜(20...     

Holographic imaging with a nanometer resolution using compact table-top EUV laser

Holographic 2D/3D imaging with nanometer resolution using short wavelength extreme ultraviolet (EUV) light is presented in this paper. Gabor’s holograms were recorded with a highly coherent table top EUV laser with different numerical apertures demonstrating ultimately a spatial resolution of 46+/−2 nm, comparable with the illumination wavelength, in 2D holographic imaging. Three dimensional images were obtained from a single high numerical aperture hologram recorded in a high resolution photoresist and numerically reconstructed at different image planes, allowing numerical optical sectioning with a lateral resolution ∼170 nm and depth resolution of 2.4 μm. The holograms were recorded in a high resolution photoresist and digitized with an atomic force microscope. To assess the spatial resolution of the numerical reconstructions of the holograms a correlation method was used. The algorithm allows for simultaneous estimation of the resolution and the feature size of the image under analysis.     

基于涡旋光照明的暗场数字全息显微方法研究

提出了一种基于涡旋光照明的暗场数字全息显微方法. 从理论上阐述了涡旋光环形照明原理和暗场数字全息显微原理, 分析了涡旋光的准无衍射特性对成像的影响; 搭建了相应的数字全息显微成像系统, 采用690 nm的聚苯乙烯小球作为实验样品; 最后通过对小球明暗场下数字全息显微再现像的分析对比, 证明该方法可以有效地提高数字全息系统的分辨率, 同时增强了再现像的对比度. 关键词： 全息 暗场数字全息显微 涡旋光 分辨率     

环型孔成像系统在部份相干光照明下对两个点物之分辨

在部份相干光照明下的环型孔成像系统，对两个点物之分辨，按照Ｓｐａｒｒｏｗ判据作了分析和研究，将两点之分辨作为两个参数的函数进行讨论－其一为照明光的空间相干条件，其二为环型孔的中心阻挡，在与圆型孔和完全相干照明的情况比较之后指出，环型孔和部份相干照明情况具有更高的两个点物的分辨本领。     

Resolution of 90 nm (lambda/5) in an optical transmission microscope with an annular condenser

Resolution of 90 nm was achieved with a research microscope simply by replacing the standard bright-field condenser with a homebuilt illumination system with a cardioid annular condenser. Diffraction gratings with 100 nm width lines as well as less than 100 nm size features of different-shaped objects were clearly visible on a calibrated microscope test slide. The resolution increase results from a known narrower diffraction pattern in coherent illumination for the annular aperture compared with the circular aperture. This explanation is supported by an excellent accord of calculated and measured diffraction patterns for a 50 nm radius disk.     

Terahertz near-field imaging using subwavelength plasmonic apertures and a quantum cascade laser source

The first demonstration, to our knowledge, of near-field imaging using subwavelength plasmonic apertures with a terahertz quantum cascade laser source is presented. "Bull's-eye" apertures, featuring subwavelength circular apertures flanked by periodic annular corrugations were created using a novel fabrication method. A fivefold increase in intensity was observed for plasmonic apertures over plain apertures of the same diameter. Detailed studies of the transmitted beam profiles were undertaken for apertures with both planarized and corrugated exit facets, with the former producing spatially uniform intensity profiles and subwavelength spatial resolution. Finally, a proof-of-concept imaging experiment is presented, where an inhomogeneous pharmaceutical drug coating is investigated.     

Resolution enhancement of random adsorbed single-molecule localization based on surface plasmon resonance illumination

Single-molecule localization (SML) is a powerful tool to overcome the diffraction limit in optical imaging, because the fluorescence emitted by single molecules can be observed with nanometer accuracy when the optical background and associated noise are made sufficiently small. Random adsorbed SML has been successfully demonstrated for superresolution imaging on metal surfaces. To optimize the random adsorbed SML, we developed a new illumination method based on surface plasmon resonance (SPR). The enhancement of the fluorescence signal and the reduction of background noise were achieved simultaneously. A high localization resolution of 15?nm was demonstrated with this new SPR illumination system.     

High-resolution beam steering using microlens arrays

Imaging or beam-steering systems employing a periodic array of microlenses or micromirrors suffer from diffraction problems resulting from the destructive interference of the beam segments produced by the array. Simple formulas are derived for beam steering with segmented apertures that do not suffer from diffraction problems because of the introduction of a moving linear phase shifter such as a prescan lens before the periodic structure. The technique substantially increases the resolution of imaging systems that employ microlens arrays or micromirror arrays. Theoretical, numerical, and experimental results demonstrating the high-resolution imaging concept using microlens arrays are presented.     

Improving Data Quality in Traditional Low-Dose Scanning Transmission Electron Microscopy Imaging

It has become very important to study and find optimal conditions for imaging electron-beam (e-beam) sensitive materials in scanning transmission electron microscopy under low electron-dose with high signal-to-noise ratio (SNR). Convergence and collection angles and electron-probe current are essential parameters. However, these parameters have rarely been discussed in a systematic way. In this paper, the illumination and collection conditions are optimized according to the resolution requirement of different materials by adjusting the condenser and intermediate lenses in a commercial transmission electron microscope. To demonstrate the significance of optimizing these parameters, two examples, zeolite MFI and metal–organic framework (MOF) MIL-101, are taken among the sensitive materials, with the most important electron incidences along the [010] and <110> directions, respectively. High SNR atomic resolution images of MFI are obtained with e-beam current as low as 0.50 pA, reaching information transfer for reflection up to 18 0 2 corresponding to d-spacing of 0.11 nm, close to the resolution limit of 0.098 nm from resolvable diffraction limit. MOF MIL-101 is characterized under an even lower e-beam 0.2 pA to avoid severe beam damage. High-quality annular dark and bright field images are obtained, which proves the wide applicability of this method on more e-beam sensitive materials.     

便携式双波段荧光眼底血管造影仪的光学设计

基于Gullstrand-Le Grand眼模型设计了一款便携式双波段眼底造影仪的光学结构。考虑人眼光学系统的像差并使用共轴式照明,设计了在主波段525和826 nm下全视场200万像素高清晰成像的光学系统,该系统可实现吲哚菁绿血管造影和荧光眼底血管造影两种方式成像。为了避免角膜中心具有大曲率而引入杂散光,照明系统采用环形光阑、中空反射镜和共轴式照明相结合的方法。为了达到系统的便携性和成像清晰性,在摄影系统中加入两个非球面结构。实验结果表明：本系统具有较大的调节能力,对-10 m-1～＋8 m-1人眼普遍适用,系统可实现30°的眼底拍摄,眼底分辨率〉108 lp/mm,畸变〈10%,达到了便携式双波段眼底造影仪的使用要求。     

非相干照明条件下的ptychographic iterative engine成像技术

在传统多波长相干衍射成像理论的基础上提出适用于 X-射线和电子束等非相干光源照明成像的改进多波长ptychographic iterative engine 方法, 同时将小孔形状和照明光谱信息用于叠代计算, 可以在非相干照明条件下精确重建出物体的强度透射像和相位透射像, 并对光源带宽对重建精度的影响进行了分析, 对于解决如何在非相干照明条件下对大尺寸物体进行精确相位成像的问题具有较好的科研和实用价值.