利用AFM和SNOM对淋巴细胞膜表面超微结构及其光学性质的初步研究

利用原子力显微镜(Atomic Force Microscopy，AFM)对淋巴细胞表面形貌进行了形态学的初步研究，观察到了其膜表面其他显微技术所不能发现的超微结构．同时也运用扫描近场光学显微镜(Scanning Near field Optical Microscopy，SNOM)对淋巴细胞进行成像，观察了其对光的透射、吸收等光学性质，并对两种成像方法进行了比较．研究发现：淋巴细胞膜表面凹凸不平，分布着大量直径几十到几百纳米不等的小颗粒；淋巴细胞中央部位有自发荧光现象．结果表明，AFM和SNOM可作为进一步探讨淋巴细胞的结构与功能关系的有力工具．     

Cantilever tip near-field surface-enhanced Raman imaging of tris(bipyridine)ruthenium(II) on silver nanoparticles-coated substrates

The near-field surface-enhanced Raman scattering (SERS) and surface-enhanced fluorescence (SEF) images of tris(bipyridine)ruthenium(II) adsorbed on a silver nanoparticles-coated substrate were obtained with a scanning near-field optical microscope (SNOM, or near-field scanning optical microscopy, NSOM) using a cantilever tip. In comparison with the most widely used fiber tip for SNOM, the cantilever tip has higher optical throughput and better thermal stability, making it more suitable for detecting the extremely low Raman signal in the near-field spectroscopic investigations. Our preliminary results show that the near-field SERS with the higher spatial resolution can provide richer fingerprint information than the far-field SERS. A comparison of the two types of images shows that there are more SERS than SEF hot spots, and the two types of hot spots do not overlap. More surprisingly, the near-field SERS spectra differ from the far-field SERS spectra obtained on the same sample in the band frequency and relative intensities of some major Raman bands, and some IR-active bands were observed with the near-field mode. These results are explained mainly by the electric field gradient effect and heterogeneous polarization character that operate only in the near-field SERS.     

扫描近场光学显微镜及其在单分子探测中的应用

扫描近场光学显微镜在光学显微镜中具有独特的性能,其突破衍射光限制,具有单分子探测灵敏度,且在研究时不损伤生物样品。文中简要介绍了扫描近场光学显微镜的原理,详述近年来扫描近场光学显微镜在单分子探测中的应用,介绍了扫描近场光学显微镜结合量子点对单分子探测的进展,并对单分子探测的前景做了展望。     

基于扫描探针显微镜(SPM)的高密度信息存储

随着信息技术的飞速发展,高密度信息存储的研究成为国际上备受关注的研究领域。扫描探针显微技术（SPM）通过改变材料的光、电、磁等局域特性可以实现纳米尺度的信息存储,成为提高信息存储密度的最有效手段之一。本文从信息存储材料和技术角度综述了基于SPM的高密度信息存储最近的研究进展,并讨论了其将来的研究和发展方向。     

Scanning near-field optical microscopy

Scanning near-field optical microscopy (SNOM) has become a widespread technique due to its promising ability of imaging with sub-micron resolution. Despite being developed over more than one decade, SNOM is still not a mature technique, which can be seen from the large number of recent publications describing instrumentational innovations. However, there are also many applications of near-field microscopy to the observation of thin organic film systems, which are supplementary to other techniques and demonstrate the usefulness of the technique.     

扫描近场光学显微镜对纳米结构的观察

光学显微镜在人们认识微观世界的过程中一直扮演着非常重要的角色．随着认识的深入，对空间分辨率的要求也越来越高．但是众所周知，普通光学显微镜（远场情况下）的分辨率受光的衍射效应所限制，一般可表达为0．61A／N．A．（约等于A／2，A为照射光的波长，N．A．为数值孔径）     

Plasmon resonances on metal tips: understanding tip-enhanced Raman scattering

Calculations of the electric-field enhancements in the vicinity of an illuminated silver tip, modeled using a Drude dielectric response, have been performed using the finite difference time domain method. Tip-induced field enhancements, of application in "apertureless" Raman scanning near-field optical microscopy (SNOM), result from the resonant excitation of plasmons on the metal tip. The sharpness of the plasmon resonance spectrum and the highly localized nature of these modes impose conditions to better exploit tip plasmons in tip-enhanced apertureless SNOM. The effect of tip-to-substrate separation and polarization on the resolution and enhancement are analyzed, with emphasis on the different field components parallel and perpendicular to the substrate.     

Electromagnetic coupling in near-field scattering by small homogeneous and heterogeneous nanoaggregates

Progress in near-field optical spectroscopy research on metal nanoparticles demands a better understanding of the role played by particle-particle interactions and a deeper insight of the influence of the incident field wavelength. This is particularly true for scanning near-field optical microscopy (SNOM), where the mechanism by which some components of the evanescent illuminating field are transformed into propagating field components that carry information about the sample is at the core of the image formation and where the role played by the interactions between sample and tip remains a still open problem. In this perspective, we investigate numerically the optical behavior of small aggregates of spherical nanoparticles, taking into account the electromagnetic coupling between all particles and the apertureless tip. The tip is modeled as a sphere made of different materials characterized by appropriate dielectric functions. We find that the tip material affects both qualitatively and quantitatively the SNOM images; more important, from the analysis of the calculated scattering cross section, the resonance plasmon location of the whole (aggregate + tip) system undergoes detectable changes, if the tip is constituted of the same material of the sample, as the tip is situated in different positions. This modification of the plasmon frequencies induces a nontrivial variation of the near-field intensity as a function of the tip position and the resulting SNOM image can be distorted with respect to the actual shape of the sample. No simple arguments can be used to relate the value of the local field on the tip surface to the scattering cross section value; depending on the tip material, the comparison between these two measurements can help to clarify the role of basic interactions in the scattering mechanism.     

Time-resolved fluorescence spectroscopic and scanning near-field optical microscopic studies of rhodamine dye adsorbed in cationic Langmuir-Blodgett films

The fluorescence lifetimes decays and picosecond time-resolved fluorescence spectra were measured to investigate the dynamics of the excited state of sulforhodamine B (SRB) molecules adsorbed in the mono- and multilayered Langmuir-Blodgett (LB) films of octadecylamine. Steady-state and time-resolved fluorescence spectroscopy reveals that the fluorescence lifetimes and contents of the monomer and dimers in the molecular organizates depend upon the concentration of the dye in the solution and the adsorption process. SRB dye molecules adsorbed in LB films have been imaged with scanning near-field optical microscopy (SNOM). This information is exploited to map the distribution with molecular spatial resolution. SNOM provide the visual evidence of the monomers and dimers of SRB in cationic LB films.     

Electrostatic interactions in protein adsorption probed by comparing lysozyme and succinylated lysozyme

In this paper, we present a nanoscale study of the supramolecular structure of the dehydrogenate polymer (ZL-DHP) lignin model compound. The combination of near-field scanning optical microscopy (NSOM or SNOM) and atomic force microscopy (AFM) has been utilized to explore physicochemical properties of the lignin model compound on a scale ranging from individual macromolecules to globular supramolecular assemblies. By utilizing NSOM in transmission mode, the optical inhomogeneity in the lignin supramolecular structure has been observed for the first time. In particular, the transmission-mode NSOM images reveal a combination of hollow and layered supramolecular globular structure in the lignin model compound. Through the paired use of TappingMode and pulsed-mode AFM, we have also confirmed the existence of regions with different rheological properties on the single lignin model compound supramolecular assembly.     

Scanning Near-Field Optical Microscopy and Spectroscopy as a Tool for Chemical Analysis

Research into the nanometer-scale region is currently of relevance in many branches of modern science and engineering, such as in microelectronics, supramolecular chemistry, and in a biological context. A great deal of attention is given to the design of molecular devices, usually towards understanding the function of existing "molecular machines". Central to this task are powerful diagnostic tools capable of recording chemical information with spatial resolution in the nanometer range. While elemental analysis of surfaces with a lateral resolution of a few dozen nanometers is almost routine, analysis of molecular species with a resolution of <1 μm is very difficult. Scanning-tunneling and atomic-force microscopies usually do not give any chemical information. By combining scanning near-field optical microscopy (SNOM)-the "optical member" of the family of scanning-probe microscopies-with optical spectroscopy, it is possible to obtain molecular information from sample areas as little as 50 nm in diameter. In SNOM, a light source is scanned above the object of interest at a distance of a few nanometers. In the optical near field, the illuminated area is not subject to the Abbé diffraction limit, but merely by the size of the illuminating source. High quality SNOM probes can be reproducibly prepared by a chemical-etching method. These etched probes have an optical transmission up to 1000-fold higher than commercial (pulled) SNOM tips and can withstand higher laser power. This last advantage allows not only high resolution optical imaging, but also localized spectroscopic investigations of surfaces and even optical "nanosampling" by pulsed-laser ablation. The ablated material can be transported over a considerable distance, which opens the possibility for its subsequent analysis with a complementary, highly sensitive analytical method, such as mass spectrometry.     

利用扫描近场光学显微镜的偏振衬度对各向异性微晶的观察

利用扫描近场光学显微镜的偏振衬度对罗丹明6G的微晶进行观察,结果表明,用圆偏振入射光进行观察时,光学图像与表面形貌图有很好的对应关系。而用线偏振入射光进行观察时,随着入射偏振态的不同,成像差别较大:当偏振态方向沿着具有各向异性吸收的晶粒主吸收方向时,在垂直于主吸收方向上有很高的衬度,但图像分辨率较低;而入射偏振态垂直于主吸收方向时,衬度降低,分辨率提高;在与主吸收方向成45°角入射时,可以得到分辨率和衬度都比较好的图像。     

Ultrafast plasmon dynamics and evanescent field distribution of reproducible surface-enhanced Raman-scattering substrates

Surface-enhanced Raman scattering (SERS) is a potent tool in bioanalytical science because the technique combines high sensitivity with molecular specificity. However, the widespread and routine use of SERS in quantitative biomedical diagnostics is limited by tight requirements on the reproducibility of the noble metal substrates used. To solve this problem, we recently introduced a novel approach to reproducible SERS substrates. In this contribution, we apply ultrafast time-resolved spectroscopy to investigate the photo-induced collective charge-carrier dynamics in such substrates, which represents the fundamental origin of the SERS mechanism. The ultrafast experiments are accompanied by scanning-near field optical microscopy and SERS experiments to correlate the appearance of plasmon dynamics with the resultant evanescent field distribution and the analytically relevant SERS enhancement. Figure Ultrafast time-resolved differential absorption spectroscopy combined with scanning near-field optical microscopy (left) and atomic force microscopy (right) yields insight into the photoinduced charge-carrier dynamics in innovative reproducible SERS-substrates Dana Cialla and Ronald Siebert contributed equally to this work.     

Sub-wavelength Raman spectroscopy on isolated silver islands

Surface Enhanced Raman Spectroscopy (SERS) is a valuable analytical tool for the investigation of molecules adsorbed on roughened noble metal surfaces. The shape, size, and surrounding of the metal protrusions play an important role in the Raman scattering enhancement. By combining scanning near-field optical microscopy (SNOM) with Raman spectroscopy the spatial resolution suffices for investigating isolated silver islands on SERS active substrates. We demonstrate an optical resolution below 70 nm for recording spectra on specifically prepared and fully characterized SERS substrates. For a quantitative evaluation of the SERS signal the spatial distribution of Rhodamine 6G (R6G) deposited on the SERS substrate was determined by friction force measurements. By comparing the Raman intensities of the SERS substrates with those of unmetallized support plates absolute SERS enhancement factors at specific locations on top and in the vicinity of the silver islands were determined directly.     

New Tools for Investigating Electromagnetic Hot Spots in Single‐Molecule Surface‐Enhanced Raman Scattering

Surface‐enhanced Raman scattering (SERS) is quickly growing as an analytical technique, because it offers both molecular specificity and excellent sensitivity. For select substrates, SERS can even be observed from single molecules, which is the ultimate limit of detection. This review describes recent developments in the field of single‐molecule SERS (SM‐SERS) with a focus on new tools for characterizing SM‐SERS‐active substrates and how they interact with single molecules on their surface. In particular, techniques that combine optical spectroscopy and microscopy with electron microscopy are described, including correlated optical and transmission electron microscopy, correlated super‐resolution imaging and scanning electron microscopy, and correlated optical microscopy and electron energy loss spectroscopy.     

单分子定位超分辨显微成像有机荧光探针的研究进展

在生物医学领域,对纳米尺寸级别的微小生物目标进行精确定位研究具有非常重要的意义,而光学显微成像技术为此提供了强有力的工具。 光学显微成像技术受到光学衍射极限的限制,难以分辨尺寸在衍射极限(<200 nm)以下的生物结构,无法直接获取微小生物结构信息,阻碍了生物医学的进一步发展。 近年来,随着纳米分辨显微成像技术的出现,新型荧光探针的开发、成像系统与设备的不断发展及成像算法不断完善地深入结合,促进了光学衍射极限以下尺寸微观目标的研究。 基于单分子定位的超分辨荧光显微成像(SMLM)包括光激活定位成像(PALM)与随机光学重构超分辨成像(STORM),将有机荧光探针与超分辨光学显微成像技术紧密结合在一起,荧光探针的光物理性质直接决定着超分辨成像结果的好坏。 因此,设计不同性能的荧光探针可以实现超精细结构的不同超分辨成像,为研究其生物学功能提供了有力的工具。 本文着重围绕基于SMLM的原理、有机荧光探针的设计要求、用于SMLM的荧光探针种类及其生物应用等方面进行总结综述,指出了单分子定位成像上存在的不足,并对其发展方向进行了展望,希望为对超分辨成像研究感兴趣或初涉该领域的研究者提供成像理论与探针设计方面的帮助。     

纳米气泡电化学研究进展

纳米气泡广泛存在于许多自然现象和工业生产过程,其自身具有独特的物理化学性质.由于涉及气体反应的纳米电催化及能源转化技术的迅速发展,有关纳米气泡的电化学研究越来越受研究者的关注.针对电极界面纳米气泡的研究不仅有助于实现对气泡行为的调控,指导催化剂电极界面的合理设计以提高电催化效率,也可以从科学上去了解纳米催化剂本征电催化...     

Tip-enhanced near-field optical microscopy

Spectroscopic methods with high spatial resolution are essential for understanding the physical and chemical properties of nanoscale materials, including quantum structures and biological surfaces. An optical technique is reviewed that relies on the enhanced electric fields in the proximity of a sharp, laser-irradiated metal tip. These fields are utilized for spatially confined probing of various optical signals, thus allowing for a detailed sample characterization far below the diffraction limit. In addition, tip-enhanced fields also provide the sensitivity crucial for the detection of nanoscale volumes. After outlining the principles of near-field optics, the mechanisms contributing to local field enhancement and how it can be used to enhance optical signals are discussed. Different experimental methods are presented and several recent examples of Raman and fluorescence microscopy with 10 nm spatial resolution of single molecules are reviewed.     

Non‐optical bimorph‐based force sensor for scanning near‐field optical microscopy of biological materials: characteristics,design and applications

A non‐optical force sensor that allows operation both in lateral (shear) and in vertical (tapping) force detection modes has been introduced for dynamic tip–sample distance regulation in scanning near‐field optical microscopy (SNOM) of biological samples. The sensor is based on a rectangular bimorph cantilever consisting of two thin piezoceramic layers bonded to a brass centre shim. One of the piezo layers serves as the probe dither and another as the responder of the sensed forces. The sensor is driven with a home‐made Q‐control electronics so that its sensitivity and bandwidth can be adjusted. The dynamics, characteristics and design considerations of the sensor are theoretically and experimentally discussed. Driving the bimorph cantilever at its eigenfrequency with appropriate force feedback allows one to obtain a quality factor (Q‐factor) up to 10³ in water, suitable for different sample softness and imaging environments. The high sensitivity of the sensor is demonstrated both by shear force and by tapping mode imaging of soft biological samples in their natural state. Near‐field optical resolution of better than 100 nm on red blood cells in water has been obtained. The experimental results suggest that this SNOM sensor would be a promising set‐up for biological applications. Copyright © 2007 John Wiley & Sons, Ltd.     

Sub-micrometer polarimetry of chiral surfaces using near-field scanning optical microscopy

In this work we have studied the optical activity of chiral crystal surfaces with polarized near-field scanning optical microscopy (NSOM); our studies clearly demonstrated that polarized NSOM can be utilized to determine chirality at crystal surfaces.