Bioimaging TOF-SIMS of tissues by gold ion bombardment of a silver-coated thin section

The imaging time-of-flight secondary-ion-mass-spectrometry (TOF-SIMS) method was utilized to address the problem of cholesterol localization in rat tissues. Rat kidneys were fixed, cryoprotected by sucrose, frozen, sectioned by cryoultramicrotomy, and dried at room temperature. The samples were either covered with a thin silver layer or analyzed uncovered in an imaging TOF-SIMS instrument equipped with an Au1-3(+)-source. The yield of desorbed secondary ions for some species was up to 600-fold higher after silver coating of the samples. Reference samples of cholesterol were silver-coated and analyzed by TOF-SIMS to define significant peaks, specific for cholesterol. Such peaks were found at m/z = 386 (C27H46O+), m/z = 493 (C27H46O107Ag+), m/z = 495 (C27H46O109Ag+), m/z = 879 (C54H92O2 107Ag+), and m/z = 881 (C54H92O2 109Ag+). The silver-cationized cholesterol (493 < or = m/z < or = 495) signal was localized by imaging TOF-SIMS in the kidney sections and showed a high cholesterol content in the kidney glomeruli. A more diffuse distribution of cholesterol was also found over areas representing the cytoplasm or plasma membrane of the epithelial cells in the proximal tubules of rat kidney.     

Bioimaging TOF-SIMS: High resolution 3D imaging of single cells

The distribution of phosphocholine ions (m/z 184, m/z 86), sodium ions, and potassium ions in thyroid tumor cells was analyzed by imaging TOF-SIMS. Repeated sputtering with a C(60) (+) source and subsequent analysis with a Bi(3) (+) gun produced a series of 138 images that were stacked to make a 3D display of the chemistry of cells. Phosphocholine was seen in the plasma membrane (m/z 184) and intracellular membranes (m/z 86). The different fragmentation of the phospholipid probably reflects the chemical composition of membranes at these sites. High intensity of secondary ion signals of potassium was seen in membrane-encompassed cellular compartments. The data indicate that potassium ions are compartmentalized in thyroid tumor cells.     

直接金属成形喷射技术

分析了现有典型直接金属成形同轴喷射喷头系统的专利,结合试验提出了喷头系统的设计准则。在此基础上设计开发了单喷点同轴喷射系统。试验证明该喷头使成形工艺控制更加灵活,提高了成形零件的质量。     

Subcellular localization of boron in cultured melanoma cells by electron energy-loss spectroscopy of freeze-dried cryosections

Boron neutron capture therapy (BNCT) is based on the ability of the non‐radioactive isotope ¹⁰B to capture thermal neutrons and to disintegrate instantaneously. This reaction opens a way to selectively destroy tumour cells after specific uptake of ¹⁰B. In this paper, a method based on electron energy‐loss spectroscopy is presented for detecting and quantifying boron in freeze‐dried cryosections of human melanoma cells. A practical detection limit of around 6 mmol kg^?1 in 0.1‐µm² areas is estimated using specimens prepared from standard boron solutions. Preliminary results of boron mapping in the spectrum‐imaging acquisition mode reveal boron penetration and probably spot‐like accumulation within melanoma cells when exposed to culture medium containing sodium borocaptate.     

Dissimilar metal deposition with a stainless steel and nickel-based alloy using wire and arc-based additive manufacturing

The wire and arc-based additive manufacturing process applies arc welding technology; the wire material is melted by the arc discharge, and is then accumulated successively in this process. The wire and arc-based additive manufacturing process directly and locally adds material to the molten pool. By changing the material locally during the process, more than one kind of material can be used simultaneously in a single manufactured component. In this study, two kinds of dissimilar metal deposition were conducted. A combination used was a stainless steel and Ni-based alloy. Mechanical properties near the interface such as hardness and bond strength were investigated. As a result, it was found that the mechanical properties of the manufactured alloy were comparable to those of a bulk material. In addition, an additive manufacturing system and a torch path planning method for using more than two kinds of material were proposed. By using this method, highly functional shapes whose surfaces and inner structures are made of different material could be made.     

基于超声振动的激光金属堆积机理研究分析

为提高316L不锈钢材料的激光堆积层硬度,提出了激光超声堆积（LUMD)方法。该方法通过基板的超声振动来改变堆积层晶粒的生长方向和生长速度,达到细化堆积层的晶粒、提高堆积层显微硬度之目的。在理论分析的基础上,通过实验测试了相同工艺参数下堆积层晶粒的大小和硬度。实验结果显示,改变LUMD的工艺参数后,堆积层晶粒得到了细化,堆积层的硬度得到了不同程度的提高。研究表明,采用LUMD方法可使堆积层的显微硬度得到显著提高。     

Cr4Mo4V轴承钢金属离子注入抗磨损、抗腐蚀性能研究

在氮气气氛环境下,采用金属等离子体浸没离子注入与沉积(MePIIID)工艺对Cr4Mo4V轴承钢表面进行强化处理,在高能钛离子轰击下,明显提高了Cr4Mo4V轴承钢近表层的抗磨损、抗腐蚀性能,最佳处理试件与未处理试件相比,摩擦系数由0.7～0.8降低到0.2～0.3之间。腐蚀电流密度减小了97%,腐蚀电阻增加近40倍,为轴承钢材料表面强化提供了一种新颖的改性方法。     

Quantitative subcellular imaging of boron compounds in individual mitotic and interphase human glioblastoma cells with imaging secondary ion mass spectrometry (SIMS)

Boron measurements at subcellular scale are essential in boron neutron capture therapy (BNCT) of cancer as the nuclear localization of boron‐10 atoms can enhance the effectiveness of killing individual tumour cells. Since tumours contain a heterogeneous population of cells in interphase as well as in the M phase (mitotic division) of the cell cycle, it is important to evaluate the subcellular distribution of boron in both phases. In this work, the secondary ion mass spectrometry (SIMS) based imaging technique of ion microscopy was used to quantitatively image boron from two BNCT agents, clinically used p‐boronophenylalanine (BPA) and 3‐[4‐(o‐carboran‐1‐yl)butyl]thymidine (N4), in mitotic metaphase and interphase human glioblastoma T98G cells. N4 belongs to a class of experimental BNCT agents, designated 3‐carboranyl thymidine analogues (3CTAs), which presumably accumulate selectively in cancer cells due to a process referred to as kinase‐mediated trapping (KMT). The cells were exposed to BPA for 1 h and N4 for 2 h. A CAMECA IMS‐3f SIMS ion microscope instrument capable of producing isotopic images with 500 nm spatial resolution was used in the study. Observations were made in cryogenically prepared fast frozen, and freeze‐fractured, freeze‐dried cells. Three discernible subcellular regions were studied: the nucleus, a characteristic mitochondria‐rich perinuclear cytoplasmic region, and the remaining cytoplasm in interphase T98G cells. In metaphase cells, the chromosomes and the cytoplasm were studied for boron localization. Intracellular concentrations of potassium and sodium also were measured in each cell in which the subcellular boron concentrations were imaged. Since the healthy cells maintain a K/Na ratio of approximately 10 due to the presence of Na‐K‐ATPase in the plasma membrane of mammalian cells, these measurements provided validation for cryogenic sample preparation and indicated the analysis healthy, well preserved cells. The BPA‐treated interphase cells revealed significantly lower concentrations of boron in the perinuclear mitochondria‐rich cytoplasmic region as compared to the remaining cytoplasm and the nucleus, which were not significantly different from each other. In contrast, the BPA‐treated metaphase cells revealed significantly lower concentration of boron in their chromosomes than cytoplasm. In addition, the cytoplasm of metaphase cells contained significantly less boron than the cytoplasm of interphase cells. These observations provide valuable information on the reduced uptake of boron from BPA in mitotic cells for BPA‐mediated BNCT. SIMS observations on N4 revealed that boron was distributed throughout the interphase and mitotic cells, including the chromosomes. The presence of boron in chromosomes of metaphase cells treated with N4 is indicative of a possible incorporation of this thymidine analogue into DNA. The 3‐D SIMS imaging approach for the analysis of mitotic cells shown in this work should be equally feasible to the evaluation of other BNCT agents.     

Electron microscopy localization and characterization of functionalized composite organic-inorganic SERS nanoparticles on leukemia cells

We demonstrate the use of electron microscopy as a powerful characterization tool to identify and locate antibody-conjugated composite organic-inorganic nanoparticle (COINs) surface enhanced Raman scattering (SERS) nanoparticles on cells. U937 leukemia cells labeled with antibody CD54-conjugated COINs were characterized in their native, hydrated state using wet scanning electron microscopy (SEM) and in their dehydrated state using high-resolution SEM. In both cases, the backscattered electron (BSE) detector was used to detect and identify the silver constituents in COINs due to its high sensitivity to atomic number variations within a specimen. The imaging and analytical capabilities in the SEM were further complemented by higher resolution transmission electron microscopy (TEM) images and scanning Auger electron spectroscopy (AES) data to give reliable and high-resolution information about nanoparticles and their binding to cell surface antigens.     

Optical in-process height measurement system for process control of laser metal-wire deposition

We propose an in-process height measurement system for a weld bead and feedback control system for wire-feeding speed for high-quality laser deposition. Metal additive manufacturing, especially laser metal-wire deposition, is effective for complex shape fabrication and repair processing. However, we must control the gap between a weld bead and a feed wire in an optimal range for high-quality deposition. Conventionally, the Z-stage pitch for multi-layer deposition must be precisely adjusted by each deposition shape. In this paper, we design an in-process height measurement system that is integrated in a laser processing head, which measures the weld bead height by a line section method. We decreased the influence of the intense thermal radiation generated from a melt pool by inserting the band-pass filter of the line beam's wavelength in the imaging system and optimizing its line laser power. Consequently, our system can measure the weld bead height near the melt pool, which is 4 mm in front of it. Next we show that our proposed system can measure the weld bead height during wire-laser metal deposition with 50-μm accuracy by comparing its value to the true value. Finally, we achieved a cylinder shape deposition of 50-mm height, regardless of the Z-stage pitch and the cylinder diameter of the multi-layer deposition, by controlling the wire-feeding speed based on the measured weld bead height.     

电纺直写纳米纤维在图案化基底的定位沉积

为进一步提高单根电纺丝纳米纤维的定位沉积和形貌控制水平,基于近场静电纺丝技术,研究了单根直写纳米纤维在无图案硅基底的沉积行为;仿真分析了图案化硅基底上方的空间电场分布;采用图案化硅基底作为收集板,实验考察了微图案形状、收集运动速度等因素对单根纳米纤维定位沉积的影响规律。实验结果显示,电纺直写技术具有良好的定位精度,可将直径为100～800 nm的纳米纤维精确定位于直径仅为1.6μm的圆形微图案阵列上表面;收集板运动速度较小时,受电场力影响纳米纤维沉积轨迹将朝微图案偏移7μm;收集板运动速度进一步减小时,纳米纤维在基底微图案附近或上表面产生聚集;长条形微图案对纳米纤维沉积过程具有良好的引导与约束作用。得到的结果表明,基于近场静电纺丝的直写技术可较好地实现单根纳米纤维在图案化硅基底的精确定位沉积。     

New kind of subarea-parallel scanning mode for laser metal deposition shaping

A scanning mode is the key technology in a laser metal deposition shaping (LMDS) fabrication process. On the basis of the analysis of existing scanning modes and their influences on the fabrication efficiency and the quality of parts, some disadvantages of them are pointed out. A new kind of subarea-parallel scanning mode for LMDS based on a subdividing profiled outline into monotonous polygon subareas is presented. First, based on the principle of point visibility, inner loops are eliminated, and simple polygons are subdivided into monotonous polygons with the minimal zones. Second, the parallel scanning paths of all monotonous polygon subareas are finished, which diminishes the length of the scanning line. The practical application shows that the scanning mode can enhance the fabrication efficiency and quality. __________ Translated from Chinese Journal of Mechanical Engineering, 2006, 42(10): 170–175 [译自: 机械工程学报]     

脉冲真空电弧离子镀发射特性的测量

通过大量工艺实验 ,镀制了改变脉冲真空电弧离子源电源参数 (主回路电压 ,脉冲频率 )、基片高度以及磁场等工艺参数的样品 ,选择了一种合适的膜厚测量仪器进行了膜厚分布测量 ,获得了离子源各种工艺参数对其发射特性影响的曲线     

脉冲激光溅射沉积技术的发展及其应用研究

薄膜材料已经在半导体材料、超导材料、生物材料等方面得到广泛应用.为了得到高质量的薄膜材料,脉冲激光溅射沉积(PLD)技术受到了广泛的关注.文中介绍了脉冲激光溅射沉积薄膜的基本原理及特点,分析了脉冲激光溅射沉积技术在制备高温超导膜、铁电薄膜、生物陶瓷薄膜等功能薄膜方面的应用研究.大量研究表明,脉冲激光溅射沉积是目前最好的...     

Fabrication of nanoscale step height structure using atomic layer deposition combined with wet etching

The current techniques used for the fabrication of nanosteps are normally done by layer growth and then ion beam thinning. There are also extra films grown on the step surfaces in order to reduce the roughness. So the whole process is time consuming. In this paper, a nanoscale step height structure is fabricated by atomic layer deposition(ALD) and wet etching techniques. According to the traceable of the step height value, the fabrication process is controllable. Because ALD technology can grow a variety of materials, aluminum oxide(Al_2O_3) is used to fabricate the nanostep. There are three steps of Al_2O_3 in this structure including 8 nm, 18 nm and 44 nm. The thickness of Al_2O_3 film and the height of the step are measured by anellipsometer. The experimental results show that the thickness of Al_2O_3 film is consistent with the height of the step. The height of the step is measured by AFM. The measurement results show that the height is related to the number of cycles of ALD and the wet etching time. The bottom and the sidewall surface roughness are related to the wet etching time. The step height is calibrated by Physikalisch-Technische Bundesanstalt(PTB) and the results were 7.5±1.5 nm, 15.5±2.0 nm and 41.8±2.1 nm, respectively. This research provides a method for the fabrication of step height at nanoscale and the nanostep fabricated is potential used for standard references.     

SiO2薄膜斜角蒸镀工艺研究

利用斜角蒸镀工艺镀制SiO2薄膜是获得低折射率薄膜的一个有效方法。通过电子束蒸发镀膜方式,利用自制的斜角蒸镀装置,研究了SiO2材料在斜角蒸镀工艺中薄膜倾斜角度与沉积角度的关系,薄膜沉积厚度与设定厚度的关系,薄膜折射率与沉积角度的关系。实验表明利用斜角蒸镀工艺镀制低折射率薄膜是可行的。实验得到了折射率为1.10的SiO2薄膜,并得到了重要的SiO2的折射率与沉积角度关系曲线。     

常压下用金属有机化学气相沉积法在HP40钢表面制备纳米氧化铝薄膜及表征

常压下用金属有机化学气相沉积法(MOCVD),以仲丁醇铝(ATSB)为前驱体、氮气为载气在HP40钢表面制备了纳米氧化铝薄膜;用光学显微镜、扫描电子显微镜及能谱仪、X射线衍射仪、原子力显微镜等研究了沉积温度等参数对氧化铝薄膜沉积速率的影响,并对其形貌进行了观察。结果表明:随着沉积温度从503K升高到713K,薄膜沉积速率从0.1mg·cm~(-2)·h~(-1)增加到0.82mg·cm~(-2)·h~(-1);当沉积温度在593～653K范围内,可获得晶粒尺寸为10～15nm的纳米氧化铝薄膜;反应的表观活化能随氮气与ATSB蒸气混合气流速增加而降低,不同的混合气流速有不同的反应级数,ATSB的反应级为0.7±0.02。     

Pressure effect of growing with electron beam-induced deposition with tungsten hexafluoride and tetraethylorthosilicate precursor

Electron beam-induced deposition (EBID) provides a simple way to fabricate submicron- or nanometer-scale structures from various elements in a scanning electron microscope (SEM). The growth rate and shape of the deposits are influenced by many factors. We have studied the growth rate and morphology of EBID-deposited nanostructures as a function of the tungsten hexafluoride (WF6) and tetraethylorthosilicate (TEOS) precursor gas pressure and growth time, and we have used Monte Carlo simulations to model the growth of tungsten and silicon oxide to elucidate the mechanisms involved in the EBID growth. The lateral radius of the deposit decreases with increasing pressure because of the enhanced vertical growth rate which limits competing lateral broadening produced by secondary and forward-scattered electrons. The morphology difference between the conical SiO(x) and the cylindrical W nanopillars is related to the difference in interaction volume between the two materials. A key parameter is the residence time of the precursor gas molecules. This is an exponential function of the surface temperature; it changes during nanopillar growth and is a function of the nanopillar material and the beam conditions.     

熔融挤压快速成形制件的质量分析

熔融沉积快速成形工艺能够使用ABS塑料、聚碳酸酯、尼龙基聚合物和其它一些材料建立高强度的、耐久的完全实用的原型。由于受工艺过程参数的影响，快速制件可能会出现表面缺陷和内部缺陷。对这些可能产生的缺陷进行分析，有效避免或减少它们的出现，提高快速制件的综合质量。     

Molecular dynamics simulation on nanotribological properties of molecular deposition film during the scan process

In this paper, a molecular dynamics simulation has been used to study the nanotribological properties of the molecular deposition film deposited on the Au (111) substrate during the scan process with an Au tip. The results show that the tilt angle and the potential energy of the molecules near the tip are both increased with decreasing distance between the tip and monolayer. Continuous slip also occurs during the slip stage. Furthermore, the regular stick-slip is in agreement with the spatial structure.