蘸笔纳米光刻术的新应用

蘸笔纳米光刻术（Dip-pen nanolithography,DPN）利用原子力显微镜（AFM）的探针把“墨水”分子传输至基底表面,使之形成自组装单分子层.DPN作为一种在物质表面构造纳米结构的技术,以其高分辨率、定位准确和直接书写等优点,在物理、化学、生物等领域的纳米尺度研究中得到了广泛应用.本文着重综述近年来DPN在纳米电路、生物芯片、化学检测、催化反应、纳米刻蚀等方面的新应用,以及它在实验研究中取得的新进展,分析了相应实验的原理,展示了这种技术的优势和发展前景.     

Nanoscale distribution of CD20 on B‐cell lymphoma tumour cells and its potential role in the clinical efficacy of rituximab

Rituximab is an exciting monoclonal antibody drug approved for treating B‐cell lymphomas and its target is the CD20 antigen which is expressed on the surface of B cells. In recent years, the variable efficacies of rituximab among different lymphoma patients have become an important clinical issue and urgently need to be solved for further development of antibodies with enhanced efficacies. In this work, atomic force microscopy (AFM) was used to investigate the nanoscale distribution of CD20 on the surface of tumour B cells from lymphoma patients to examine its potential role in the clinical therapeutic effects of rituximab. By performing ROR1 fluorescence labelling (ROR1 is a specific tumour cell surface marker) on the bone marrow cells prepared from B‐cell lymphoma patients, the tumour B cells were recognized, and then AFM tips carrying rituximabs via polyethylene glycol crosslinkers were moved to the tumour cells to probe the specific CD20‐rituximab interactions. By applying AFM single‐molecule force spectroscopy (SMFS) at the local areas (500×500 nm²) on the surface of tumour B cells, the nanoscale distributions of CD20 on the surface of tumour B cells were mapped, visually showing that CD20 distributed heterogeneously on the cell surface. Bone marrow cell samples from three clinical B‐cell lymphoma cases were collected to analyze the binding affinity and nanoscale distribution of CD20 on tumour cells. The experimental results showed that CD20 distribution on tumour cells were to some extent related to the clinical therapeutic outcomes while the CD20‐rituximab binding forces did not have distinct effects to the clinical outcomes. These results can provide novel insights in understanding the rituximab's clinical efficacies from the nanoscale distribution of CD20 on the tumour cells at single‐cell and single‐molecule levels.     

Speed‐ and topography‐dependent boundary friction characteristics of steel

A novel tribometer that undergoes significant changes at ultra‐slow (>5.0 µm s⁻¹) to moderate (<20 cm s⁻¹) sliding speeds was developed in order to study the friction‐speed characteristics of steel lubricated with oil. Three different surface topographies were applied to the specimens, and the friction characteristics with an additive‐free base oil (MO91) and a stearic acid‐formulated oil (StA/MO91) were studied to understand the effects of surface textures on the lubrication performance of an adsorbed molecular layer formed by StA. Friction reduction behaviour of the adsorbed layer observed in the transverse direction was attributed to microscopic hydrodynamic action that maintained the load‐carrying performance of the adsorbed layer. Copyright © 2010 John Wiley & Sons, Ltd.     

Histometric and histomorphologic comparison of combustion and ambustion using in vivo reflectance‐confocal microscopy

Background: When combustion and ambustion induce a superficial injury, they are summarized as superficial burns, regardless of the underlying cause. Reflectance‐confocal microscopy (RCM) allows noninvasive imaging of the human skin on morphological features. We hypothesized that combustion and ambustion have different histomorphological effects on the human skin. Methods: Superficial burns caused by combustion (CO‐group, five females, three males; aged 26.8 ± 14.2 years) and caused by ambustion (AM‐group, four females, four males; aged 28.1 ± 13.8 years) were evaluated 24 h after injury. The following parameters were obtained using RCM on injured and noninjured (control) site: horny layer thickness, epidermal thickness, granular cell size, basal layer thickness. Results: Compared with the controls (12.8 ± 2.5 μm), horny layer thickness decreased significantly to 10.6 ± 2.1 μm in the CO‐group, whereas it increased significantly to 17.8 ± 2.8 μm in the AM‐group. The epidermal thickness did not differ significantly in CO‐group (47.9 ± 2.1 μm) and AM‐group (49.0 ± 3.1 μm), however, both increased significantly compared with the controls (42.7 ± 1.6 μm). The basal layer thickness increased more in AM‐group (17.0 ± 1.2 μm) compared to CO‐group (15.4 ± 1.1 μm). Both differed significantly compared with their controls (13.9 ± 0.9 μm). The granular cell size increased significantly in both groups ompared to the controls (721 ± 42 μm), however, a significantly higher increase was observed in CO‐group compared to AM‐group (871 ± 55 μm vs. 831 ± 51 μm). Conclusions: RCM evaluates significant histomorphological differences in superficial burns caused by combustion and ambustion. The term “superficial burn” should consider the underlying cause and thus supplemented by the term “combustion” or “ambustion.” Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

Micro‐fabrication and monitoring of three‐dimensional microstructures based on laser‐induced thermoplastic formation

This article reports a novel laser‐induced micro‐fabrication method and its monitoring system for three‐dimensional (3D) microstructures. The mechanism of the method is that a small zone of thermoplastic material melted by laser heating grows in liquid surrounding environment, solidifying into a convex microstructure, such as micro‐dot or micro‐pillar. A laser diode (808 nm) with maximum power output of 130 mW is used as power source, and a kind of paraffin mixed with stearic acid and paint serves as the thermoplastic material for 3D microstructure formation experiments. A light microscope system consisting of a charge‐coupled device (CCD) and a computer is utilized to realize real‐time observation of the micro‐fabricating process. The distribution of local temperature rise on material surface created by laser irradiation is simulated. The effects of liquid environment on microstructure formation have been theoretically analyzed and experimentally studied. Experiments are further carried out to investigate the relationship between laser spot and fabricated microstructures. The results indicate that the widths of micro‐dots or micro‐pillars are mostly determined by the size of focal spot, and their heights increase with the enlargement of laser power density. With this method, a micro‐dot array of Chinese characters meaning “China” has been successfully fabricated through computer programming. This method has the advantages of implementing direct, mask‐less, real‐time and inexpensive 3D microstructure fabrication. Therefore, it would be widely applied in the fields of micro/nano‐technology for practical fabrication of different kinds of 3D microstructures. Microsc. Res. Tech., 2009. © 2009 Wiley‐Liss, Inc.     

Scanning electron microscopy comparison of the resin–dentin interface using different specimen preparation methods

Microscopy has been widely used to complement the data of studies related to dentin bonding; however, different specimen preparation methods may influence the analysis. Aiming to contribute to the reported scenario, this study evaluated the effect of two specimen‐sectioning methods (cleavage and diamond disk cut) on the quality of the scanning electron microscopy (SEM) images. Four crowns of human molars were selected and had an area of approximately 6 mm² of dentin exposed. They were then divided into two groups according to the universal adhesive application: total and self‐etching modes. Then, composite blocks were built up and the specimens were stored in deionized water to allow the postcuring. The specimens were further divided according to the sectioning method: cleavage or diamond disk cut. Four specimens were obtained from each tooth. They were desiccated, placed on aluminum stubs, sputter‐coated with gold, and observed in a scanning electron microscope, with ×2000 of magnification. The quality of the SEM images were evaluated by two calibrated examiners and classified into four scores (1–4). Mann–Whitney test (p < .05) showed that the diamond disk provided significantly higher scores than cleavage, whereas no significant difference was observed when comparing the total‐etching and self‐etching modes of application. The diamond disk cut method is preferable to the cleavage method to ensure the quality of the SEM analysis in studies involving the resin–dentin interface.     

单晶硅料摆辅助多金刚线切片的锯切力模型研究

料摆辅助多金刚线切片技术是实现硬脆材料高精高效加工行之有效的工艺技术,探明其工艺参数、锯切力和切片质量的定量关系,具有重要的现实意义。在研究金刚线运动轨迹的基础上,推导了考虑线弓影响的切割长度变化公式;结合压痕断裂力学和试验研究,建立并验证了料摆辅助切片的锯切力模型。开展了不同工艺参数对锯切力的影响分析,结果表明,料摆辅助加工可以降低锯切力近50%;摆动角度对最大切割力的影响较小,但摆角增大会加剧"锯齿形波动"周期内的锯切力极值幅度,摆动角速度对"锯齿形波动"的周期影响较大;在恒定进给速度条件下,进给速度越高,锯切力越大;在变速进给条件下,最大锯切力可降低12%左右。进一步进行了摆角分别为0°、3°、5°和7°的多金刚线切割单晶硅实验,试验表明,料摆辅助切片加工有助于减少硅片表面因脆性崩裂产生的表面材料破损、深凹坑等缺陷;相较于普通切片加工,在摆角5°工况时,工件的表面粗糙度和硬化层厚度最大分别降低30.1%和20.1%。     

Construction of phase diagram up to 2 GPa and 200°C for thermo‐reversible gel lubricants by diamond anvil cell

The objective of this research was to make the phase diagram up to 2.0 GPa and 200°C of thermo‐reversible gel lubricants (TR Gel‐Lube) using diamond anvil cell (DAC). TR Gel‐Lube consisted of base fluid and gelling agent. The most important characteristic of TR Gel‐Lube is the reversible change of its physical state. The DAC is able to concerning visual observation of the sol/gel and liquid/viscoelastic–solid transition at high pressure and high temperature. DAC enables the visual observation of the photoelastic effect in order to generate local principal stress difference under the compression field for liquid/viscoelastic‐solid transition of base oil poly‐α‐olefin. The direct observation for sol/gel transition of TR Gel‐Lube at high pressure was also made possible. The phase diagram corresponding to transition from liquid to viscoelastic solid for base oil poly‐α‐olefin and sol to gel for TR Gel‐Lube was made by using DAC. Copyright © 2009 John Wiley & Sons, Ltd.     

Micromorphology analysis and bond strength of two adhesives to Er,Cr:YSGG laser‐prepared vs. Bur‐prepared fluorosed enamel

Preservation of enamel during composite veneer restorations of fluorosed teeth could be achieved by conservative preparation with Erbium lasers. This study evaluated the effect of fluorosed enamel preparation with Er,Cr:YSGG vs. conventional diamond bur on the micromorphology and bond strength of a self‐etch and an etch‐and‐rinse adhesives. Er,Cr:YSGG laser or diamond bur preparation was performed on the flattened midbuccal surfaces of 70 extracted human premolars with moderate fluorosis (according to Thylstrup and Fejerskov index, TFI = 4–6). Adper Single Bond (SB) with acid etching for 20 or 40 s and Clearfil SE Bond (SEB) alone or with additional etching was applied in four laser groups. The same adhesive procedures were used in three bur groups except for 40 s of etching along with SB. After restoration, microshear bond strength was measured (MPa). Data were analyzed using ANOVA and Tamhane tests (α = 0.05). Six additional specimens were differently prepared and conditioned for scanning electron microscopy evaluation. The highest and lowest bond strengths were obtained for bur‐prepared/SB (39.5) and laser‐prepared/SEB (16.9), respectively, with a significant difference (P = 0.001). The different adhesive procedures used associated to two adhesives exhibited insignificantly lower bonding in laser‐prepared groups compared to bur‐prepared ones (P > 0.05), with the exception of additional etching/SEB, which bonded significantly higher to bur‐prepared (36.4) than to laser‐prepared enamel (18.7, P = 0.04). Morphological analyses revealed a delicate etch pattern with exposed enamel prisms on laser‐prepared fluorosed enamel after acid etching and less microretentive pattern after self‐etching primer. The etch‐and‐rinse adhesive was preferred in the laser‐prepared fluorosed enamel in terms of bonding performance. Microsc. Res. Tech. 77:779–784, 2014. © 2014 Wiley Periodicals, Inc.     

Tribological characterisation of hard coatings with and without DLC top layer in fretting tests

The potential of coatings to protect components against wear and to reduce friction has led to a large variety of protective coatings. In order to check the success of coating modifications and to find solutions for different purposes, initial tests with laboratory tribometers are usually done to give information about the performance of a coating. Different Ti‐based coatings (TiN, Ti(C,N), and TiAlN) and NiP were tested in comparison to coatings with an additional diamond‐like carbon (DLC) top coating. Tests were done in laboratory air at room temperature with oscillating sliding (gross slip fretting) with a ball‐on‐disc arrangement against a ceramic ball (Al₂O₃). Special attention was paid to possible effects of moisture (relative humidity). The coefficient of friction was measured on line, and the volumetric wear at the disc was determined after the test from microscopic measurements of the wear scar and additional profiles. The friction and wear behaviour is quite different for the different coatings and depends more or less on the relative humidity. The DLC coating on top of the other coatings reduces friction and wear considerably. In normal and in moist air the coefficient of wear of the DLC top‐layer coating is significantly less than 10⁻⁶ mm³/Nm and the coefficient of friction is below 0.1. In dry air, however, there is a certain tendency to high wear and high friction. Copyright © 2006 John Wiley & Sons, Ltd.     

Accurate and precise determination of isotopic ratios by MC‐ICP‐MS: A review

For many decades the accurate and precise determination of isotope ratios has remained a very strong interest to many researchers due to its important applications in earth, environmental, biological, archeological, and medical sciences. Traditionally, thermal ionization mass spectrometry (TIMS) has been the technique of choice for achieving the highest accuracy and precision. However, recent developments in multi‐collector inductively coupled plasma mass spectrometry (MC‐ICP‐MS) have brought a new dimension to this field. In addition to its simple and robust sample introduction, high sample throughput, and high mass resolution, the flat‐topped peaks generated by this technique provide for accurate and precise determination of isotope ratios with precision reaching 0.001%, comparable to that achieved with TIMS. These features, in combination with the ability of the ICP source to ionize nearly all elements in the periodic table, have resulted in an increased use of MC‐ICP‐MS for such measurements in various sample matrices. To determine accurate and precise isotope ratios with MC‐ICP‐MS, utmost care must be exercised during sample preparation, optimization of the instrument, and mass bias corrections. Unfortunately, there are inconsistencies and errors evident in many MC‐ICP‐MS publications, including errors in mass bias correction models. This review examines “state‐of‐the‐art” methodologies presented in the literature for achievement of precise and accurate determinations of isotope ratios by MC‐ICP‐MS. Some general rules for such accurate and precise measurements are suggested, and calculations of combined uncertainty of the data using a few common mass bias correction models are outlined. © 2009 Crown in the right of Canada. Published by Wiley Periodicals, Inc., Mass Spec Rev 28:990–1011, 2009     

Combining Ar ion milling with FIB lift-out techniques to prepare high quality site-specific TEM samples

Focused ion beam (FIB) techniques can prepare site‐specific transmission electron microscopy (TEM) cross‐section samples very quickly but they suffer from beam damage by the high energy Ga⁺ ion beam. An amorphous layer about 20–30 nm thick on each side of the TEM lamella and the supporting carbon film makes FIB‐prepared samples inferior to the traditional Ar⁺ thinned samples for some investigations such as high resolution transmission electron microscopy (HRTEM) and electron energy loss spectroscopy (EELS). We have developed techniques to combine broad argon ion milling with focused ion beam lift‐out methods to prepare high‐quality site‐specific TEM cross‐section samples. Site‐specific TEM cross‐sections were prepared by FIB and lifted out using a Narishige micromanipulator onto a half copper‐grid coated with carbon film. Pt deposition by FIB was used to bond the lamellae to the Cu grid, then the coating carbon film was removed and the sample on the bare Cu grid was polished by the usual broad beam Ar⁺ milling. By doing so, the thickness of the surface amorphous layers is reduced substantially and the sample quality for TEM observation is as good as the traditional Ar⁺ milled samples.     

Chemical structure of phosphazenes in relation to the tribological properties of a steel‐on‐steel system

The tribological behaviours of a steel‐on‐steel contact lubricated by three different fluids: (i) tetrakis(3‐trifluoromethylphenoxy)‐bis(4‐fluorophenoxy)‐cyclotriphosphazene (X‐1P), (ii) a synthetic bridged cyclotriphosphazene, and (iii) linear phosphazene derivative as base fluids were comparatively investigated on an Optimol SRV oscillating friction and wear tester. The results show that X‐1P records the lowest friction coefficient, and linear phosphazene oligomer gives the lowest wear volume loss of the steel among the investigated lubricants. Moreover, the bridged cyclotriphosphazene shows much better anti‐wear ability than cyclotriphosphazene (X‐1P) at both room and elevated temperatures. The worn surfaces were analysed by means of scanning electron microscopy, energy dispersive spectrometry and X‐ray photoelectron spectroscopy. The results demonstrate that the protective layer originated from the tribochemical reaction together with the adsorbed boundary lubricating layer containing organic F‐containing compounds; nitrogen oxide and FeF₂ played an important role in improving the friction and wear behavior of the steel‐on‐steel system. Copyright © 2009 John Wiley & Sons, Ltd.     

Differences between nanoscale structural and electrical properties of AZO:N and AZO used in polymer light‐emitting diodes

Conducting atomic force microscopy and scanning surface potential microscopy were adopted to investigate the nanoscale surface electrical properties of N‐doped aluminum zinc oxide (AZO:N) films that were prepared by pulsed laser deposition (PLD) at various substrate temperatures. Experimental results demonstrated that when the substrate temperature is 150°C and the N₂O background pressure is 150 mTorr, the N‐dopant concentration on the surface is optimal. In addition, the root‐mean‐square roughness value of the film surface, the low contact current (<400 nA) conducting region as a percentage of the total area, and the mean work function value are 1.43 nm, 96.9%, and 4.88 eV, respectively, all of which are better than those of the optimal AZO film made by PLD. This result indicates that N‐doped AZO films are better for use as window materials in polymer light‐emitting diodes. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

Immunohistochemical characterization of TH13‐L2 spinal ganglia neurons in sheep (Ovis aries)

Spinal ganglia (SG) neurons are commonly classified according to various specific features. The most widespread classification based on morphological and ultrastructural features subdivides SG neurons into light and small dark neurons. Using immunohistochemical, histochemical and lectin methods, it is possible to further subdivide the small dark neurons into two subpopulations: peptidergic and nonpeptidergic neurons. The majority of studies on SG neurons were carried out on mice and rats; there are few or no studies on large mammals. In this study, some of the widely used neuronal markers, neurofilament 200 kDa (NF200), substance P (SP), calcitonin gene‐related peptide (CGRP) and isolectin B4 (IB4), were employed to characterize neuronal nitric oxide synthase (nNOS)‐immunoreactivity (‐IR) in sheep (Ovis aries) SG (Th13‐L2) neurons. The majority of the SG neurons were IB4‐labeled (79 ± 10%), followed by NF200‐ (45 ± 4%), NOS‐ (44 ± 10%), SP‐ (42 ± 5%) and CGRP‐IR (35 ± 7%) neurons. The triple staining experiments showed that a higher percentage (75 ± 16%) of NOS‐IR neurons bound both IB4 and CGRP, or both IB4 and SP (49 ± 6%). The IB4 marker showed an unexpected staining pattern; in fact, IB4‐labeled neurons largely colocalized with NF200, usually considered a marker of light SG neurons, and with CGRP and SP. For this reason, IB4 cannot be employed in sheep to differentiate between light and dark neurons, or between peptidergic and nonpeptidergic neurons. These results suggest the importance of being cautious when comparing data among different species. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

Fabrication of polycrystalline diamond microtool using a Q-switched Nd:YAG laser

A Q-switched Nd:YAG laser (1,064 nm, 100 ns) was used to machine 2?×?1.5?×?0.5-mm rhombus-shaped tool inserts from a 60?×?0.5-mm circular disk of polycrystalline diamond. A systematic experimental study was undertaken to examine the effects of pulse repetition rate, feed rate, and number of laser passes on kerf, material removal rate, recast layer, surface morphology, and surface roughness. The optimal laser parameters for generating two-dimensional tool profiles were an average power of 3 W, a pulse repetition rate of 2 kHz, a feed rate of 1 mm/s, and a total of 45 laser passes. The beneficial results were a material removal rate of 0.02 mm³/min, kerf width of 27 μm, cutting edge radius of 6 μm, and surface roughness (Ra) of 0.625 μm. Recast layer formation, undulations, and striations were observed in the laser-cut regions. These features were attributed to the presence of a molten layer of cobalt binder, and amorphous carbon and graphite transitioned from diamond. An intriguing feature is the presence of fine particulate matter ranging in size from nanometers to a few micrometers in the laser-cut regions. It is believed that phase transition of diamond and cobalt during laser machining created thermal expansion mismatch stresses sufficient to fracture the solid into fine fragments.     

Advances in analysis of microbial metabolic fluxes via 13C isotopic labeling

Metabolic flux analysis via ¹³C labeling (¹³C MFA) quantitatively tracks metabolic pathway activity and determines overall enzymatic function in cells. Three core techniques are necessary for ¹³C MFA: (1) a steady state cell culture in a defined medium with labeled‐carbon substrates; (2) precise measurements of the labeling pattern of targeted metabolites; and (3) evaluation of the data sets obtained from mass spectrometry measurements with a computer model to calculate the metabolic fluxes. In this review, we summarize recent advances in the ¹³C‐flux analysis technologies, including mini‐bioreactor usage for tracer experiments, isotopomer analysis of metabolites via high resolution mass spectrometry (such as GC‐MS, LC‐MS, or FT‐ICR), high performance and large‐scale isotopomer modeling programs for flux analysis, and the integration of fluxomics with other functional genomics studies. It will be shown that there is a significant value for ¹³C‐based metabolic flux analysis in many biological research fields. © 2009 Wiley Periodicals, Inc., Mass Spec Rev 28:362–375, 2009     

X‐ray absorption near‐edge structure analysis of tribofilms generated by highly efficient,friction‐reducing additive in water–polyglycol–glycol lubricant

The development of environment‐friendly lubricant additives can make a valuable contribution to addressing human health and energy problems. In the present work, the compound ammonium tetrathiomolybdate was synthesised and used as a highly efficient, friction‐reducing additive in water–polyglycol–glycol lubricants. Its tribological properties were tested by a four‐ball machine, and the composition of the tribofilm was identified by X‐ray absorption near‐edge structure (XANES) spectroscopy. The tribological test results indicated that the additive cannot only reduce the friction coefficient value, but can also shorten the rubbing time needed to produce a ‘stable friction coefficient value’. The XANES results showed that the tribofilm is mainly composed of an adsorbed layer and a tribochemical layer; the formation of sulphate and molybdenum trioxide in the tribofilm on the metal wear surface is responsible for the excellent anti‐friction performances of the additive. Copyright © 2009 John Wiley & Sons, Ltd.     

Dip Pen Nanolithography: a "Desktop Nanofab" approach using high-throughput flexible nanopatterning

The ability to perform controllable nanopatterning with a broad range of "inks" at ambient conditions is a key aspect of the dip pen nanolithography (DPN) technique. The traditional ink system to demonstrate DPN is n-alkanethiols on a gold substrate, but the DPN method has found numerous other applications since. This article is meant to outline recent advances in the DPN toolkit, both in terms of research and patterning technology, and to discuss applications of DPN as a viable nanofabrication method. We will summarize new DPN developments, and introduce our concept of the "Desktop Nanofab." In addition, we outline our efforts to commercialize DPN as a viable nanofabrication technique by demonstrating massively parallel nanopatterning with the 55,000 tip 2D nano PrintArray. This demonstrates our ability to overcome the serial nature of DPN patterning and enable high-throughput nanofabrication.