Detection of protein microarrays by oblique-incidence reflectivity difference technique

Biological microarrays with different proteins and different protein concentrations are detected without external labeling by an oblique-incidence reflectivity difference (OIRD) technique. The initial experiment results reveal that the intensities of OIRD signals can distinguish the different proteins and concentrations of protein. The OIRD technique promises feasible applications to life sciences for label-free and high-throughput detection.     

Parallel detection and quantitative analysis of specific binding of proteins by oblique-incidence reflectivity difference technique in label-free format

In this work, we parallelly detected the specific binding between microarray targets including 12 different kinds of proteins and the probe solution containing five corresponding antibodies and quantitatively analyzed the interactions between CDH13 and solution phase anti-CDH13 at six different probe concentrations by oblique-incidence reflectivity difference (OIRD) method in label-free format. The detection sensitivity reached 10 ng/mL. The experimental results indicate that the OIRD method is a promising and competing technique not only in research work but also in clinic.     

用斜入射光反射差法无标记实时监测不同浓度兔IgG和山羊抗兔IgG反应的动力学过程

用斜入射光反射差法无标记实时监测1.25 mg/mL, 2.50 mg/mL和5.00 mg/mL三种不同浓度兔IgG和浓度为0.02 mg/mL山羊抗兔IgG反应的动态过程, 同时监测和获得三种不同浓度IgG反应的动力学曲线以及相应的反应时间. 实验结果表明, 斜入射光反射差法不仅能无标记地判别生物分子之间是否发生反应, 而且能无标记实时监测生物分子反应的动力学过程, 在生物分子相互作用的研究方面具有潜在和广泛的应用前景.     

Real-time and label-free detection of biomolecular interactions by oblique-incidence reflectivity difference method

We successfully conduct the label-free and real-time detection of the interactions between epoxy groups and rabbit IgG and 5' CTT CAG GTC ATG AGC CTG AT 3' oligonucleotide, and between the hybridization of 5' CTT CAG GTC ATG AGC CTG AT 3' and its complementary 3' GAA GTC CAG TAC TCG GAC TA 5' oligonucleotide, by the oblique-incidence reflectivity difference (OI-RD) method. The dynamic curves of OI-RD signals, corresponding to the kinetic processes of biomolecular combination or hybridization, are acquired. In our case, the combination of epoxy groups with rabbit IgG and 5' CTT CAG GTC ATG AGC CTG AT 3' oligonucleotide need almost one and a half hours and about two hundred seconds, respectively; and the hybridization of the two oligonucleotides needs about five hundred seconds. The experimental results show that the OI-RD is a promising method for the real-time and label-free detection of biomolecular interactions.     

Label-free detection of hybridization of oligonucleotides by oblique-incidence reflectivity difference method

The microarrays of 20-base oligonucleotide with different concentrations are detected before and after hybridization by the oblique-incidence reflectivity difference (OI-RD) method. The experimental results prove that OI-RD is a label-free method which can not only distinguish the concentration difference of oligonucleotides before and after the hybridization but also detect the hybridization of short oligonucleotides. At present the OI-RD method can detect 0.39 μmol/L 20-base oligonucleotide or less. These results suggest that the OI-RD method is a promising and potential technique for label-free detection of biological microarrays.     

Label-free detection of microarrays of biomolecules by oblique-incidence reflectivity difference microscopy

We developed an oblique-incidence reflectivity difference (OI-RD) scanning microscope for label-free imaging of microarrays of biomolecules upon solid substrates. We demonstrate that hybridization reactions in an oligonucleotide microarray fabricated upon a glass slide can be detected by such an OI-RD microscope.     

Detection of hybridization of protein microarrays using an oblique-incidence reflectivity difference method

Mouse-Immunoglobulin G(mouse-IgG) with different concentrations in a range from 1000 to 0.0128 μg/mL and a specific hybridization with goat anti-mouse IgG were detected successfully by using an oblique-incidence reflectivity difference(OI-RD) method.Two detection signals,consisting of an imaginary part(Im{Δp-Δs}) and a real part(Re{Δp-Δs}) of OI-RD,were obtained simultaneously.The detection results of hybridization by OI-RD were in accord with that of traditional fluorescent scans.In particular,we label-freely detected the washed mouse-IgG microarray with a series of concentrations and acquired a linear correlation between OI-RD intensities and the protein concentrations in logarithmic coordinates.The detection sensitivity of OI-RD can reach 14 fg.These experimental results suggest that the OI-RD method has potential applications in proteomics and clinical diagnosis.     

Detection of microarray protein biomolecules by oblique-incidence reflectivity difference technique without labelling agents

This paper reports that the detection to the protein in microarray format is carried out by oblique-incidence reflectivity difference （OI-RD） analysis without any labelling agents. The OI-RD intensities not only depend on the protein structure, but also vary with the protein concentration. The results indicate that this method should have potential application in detection of biochemical processes. The high throughout and in situ detection can be achieved by this method with further improving of the experimental system.     

用光腔衰荡技术测量镜片的反射率

根据光腔衰荡光谱技术原理,建立了测量镜片反射率的实验装置。利用该装置测定了一对反射率相等的高反射腔镜,反射率测试结果为（99.925±0.001）%;以22.5°将直腔变为折叠腔,测得的反射率为（99.992±0.003）％。重复测定反射镜样品的反射率,精度达到10－5。该测量装置可用于超低损耗薄膜高反射镜反射率的精确测量。轻微移动探测器位置,对腔镜的测试结果影响不大。     

一种测量光学元件高反射（透过）率的有效方法

本文提出一种测量激光谐振腔反射镜高反射率和测量镀有减反膜的高透过率光学元件的有效方法.特点是同时测量几块高反射率凹面镜或高透过率光学元件,其误差是普通单块测量方法的1/n.详细分析了一组激光腔镜高反射率和一组镀减反膜的平行薄片高透过率的测量结果.     

A simple solution to systematic errors in density determination by X-ray reflectivity: The XRR-density evaluation (XRR-DE) method

X-ray reflectivity (XRR) is a non-destructive, accurate and fast technique for evaluating film density. Indeed, sample-goniometer alignment is a critical experimental factor and the overriding error source in XRR density determination. With commercial single-wavelength X-ray reflectometers, alignment is difficult to control and strongly depends on the operator. In the present work, the contribution of misalignment on density evaluation error is discussed, and a novel procedure (named XRR-density evaluation or XRR-DE method) to minimize the problem will be presented. The method allows to overcome the alignment step through the extrapolation of the correct density value from appropriate non-specular XRR data sets. This procedure is operator independent and suitable for commercial single-wavelength X-ray reflectometers. To test the XRR-DE method, single crystals of TiO₂ and SrTiO₃ were used. In both cases the determined densities differed from the nominal ones less than 5.5%. Thus, the XRR-DE method can be successfully applied to evaluate the density of thin films for which only optical reflectivity is today used. The advantage is that this method can be considered thickness independent.     

Measurement of inner surface roughness of capillary by an x-ray reflectivity method

The inner surface roughness of a capillary is investigated by the reflectivity of x-rays penetrating through the capillary. The results are consistent with the data from atomic force microscope (AFM). The roughness measured by this new method can reach the order of angstroms with high quality capillaries.     

反射率测量中的热辐射影响及其消除方法

 反射率测量过程中,被激光加热的材料发出热辐射光,对探测器测量存在明显干扰,给测量结果带来了较大误差,该现象普遍存在于以往的测试工作中。通过深入分析热辐射特性,在反射率测量装置中加入透过波长不同于加载激光波长的探测器,探测热辐射在该透过波长下的光强信号,并利用比对法消除了反射率测量过程中的热辐射干扰,使高温段的反射率测量精度得到较大提高。     

反射率测量中的热辐射影响及其消除方法

反射率测量过程中,被激光加热的材料发出热辐射光,对探测器测量存在明显干扰,给测量结果带来了较大误差,该现象普遍存在于以往的测试工作中。通过深入分析热辐射特性,在反射率测量装置中加入透过波长不同于加载激光波长的探测器,探测热辐射在该透过波长下的光强信号,并利用比对法消除了反射率测量过程中的热辐射干扰,使高温段的反射率测量精度得到较大提高。     

Laser diagnostics of nanoscale dielectric films on absorbing substrate by differential reflectivity and ellipsometry

The differential reflection characteristics for ultrathin inhomogeneous dielectric film on absorbing substrate are investigated in the long-wavelength approximation. The obtained first-order expressions for differential reflectivity and changes in the ellipsometric angles caused by ultrathin layer are of immediate interest to the solution of the inverse problem. The method to determine the averaged values (not the realistic profile) of refractive index for inhomogeneous nanometric films are shown. The novel possibilities for determining the dielectric constant and thickness of nanoscale homogeneous films by the differential ellipsometric and reflectivity measurements are developed, and a simple method to estimate whether the nanometric film is homogeneous or not is also discussed.     

Laser diagnostics of nanoscale dielectric films on transparent substrate by integrating differential reflectivity and ellipsometry

The effect of nanometer dielectric films on the differential reflection characteristics of linearly polarized light from non-absorbing materials is investigated in the long-wavelength approximation. The second-order formulas for changes in the reflectance of s- and p-polarized light caused by ultrathin layer are obtained. A detailed analysis of the influence of ultrathin film to the reflectivity of p-polarized light in the vicinity of the Brewster angle is carried out. The novel methods are developed for determining the thickness and refractive index of uniform (or the average values of refractive index of nonuniform) nanometer-scale films by differential reflectivity and ellipsometric measurements.     

The effect of relaxation on the epitope mapping by saturation transfer difference NMR

The effect of longitudinal relaxation of ligand protons on saturation transfer difference (STD) was investigated by using a known binding system, dihydrofolate reductase and trimethoprim. The results indicate that T1 relaxation of ligand protons has a severe interference on the epitope map derived from a STD measurement. When the T1s of individual ligand protons are distinctly different, STD experiments may not give an accurate epitope map for the ligand-target interactions. Measuring the relaxation times prior to mapping is strongly advised. A saturation time shorter than T1s is suggested for improving the potential epitope map. Reduction in temperature was seen to enhance the saturation efficiency in small to medium size targets.     

Nanometric thinning of bonded silicon wafers using sacrificial anodic oxidation and investigated by X-ray reflectivity

X-ray reflectivity and atomic force microscopy are used to investigate silicon oxide ultra-thin films. Quantitative results are shown using a reflectivity simulation model based on kinematical X-ray theory. Changes in film thickness are discussed in relation to current density, voltage, charge and anodization time. The density and resistivity of silicon oxide are calculated and compared to that of thermal oxide. The electrical field existing in the layer during anodization is estimated. Surface roughness is also measured locally and averaged over the entire surface, producing a low value that meets microelectronic requirements. Thickness is carefully controlled. We show that ultra-thin silicon oxide films are of very high quality. Similar investigations are made on a twisted bonded silicon substrate obtained by the molecular bonding of two silicon wafers. It is shown that the silicon oxide is also of very good quality and can be used as a sacrificial silicon oxide in thinning down the upper silicon film. Controlled, accurate thinning is achieved down to a thickness of 10 nm, the level which is required for etching the dislocation network present at the bonding interface.     

对差分光学吸收光谱法的监测数据进行实时预测研究

差分光学吸收光谱法（DOAS）已经成为测量大气中微量气体浓度广泛应用的方法.通过对大气差分吸收光谱的分析可以得到它们的浓度.但在恶劣气候条件下，DOAS系统不能获得连续的实时监测数据，因此不能满足我国环境监测条例对环境监测子站的要求.文中提出了一种利用逐步回归分析的方法，对DOAS系统的监测数据进行实时预测.通过大量的对比实验表明，利用该方法得到的预测结果与实际测量结果一致，目前该技术已经被应用于DOAS监测仪器中. 关键词： 差分光学吸收光谱法（DOAS） 逐步回归分析 预测     

共孔径偏振耦合分光系统中反射镜造成的相位延迟差的测量

 激光信标共孔径发射接收偏振分光系统的动态相位补偿需要测量望远镜各反射镜对s光和p光的相位延迟差。利用Stokes矢量和Mueller矩阵建立了物理模型,并推导出用测得的回光功率计算相位延迟差的解析式。提出一种通过实验测量回光功率计算反射镜相位延迟差的方法,解决了在0～2p范围内唯一确定反射镜相位延迟差的问题。实际测量了两块反射镜的相位延迟差,并将测量结果用于动态相位补偿偏振分光实验,验证了该方法的正确性。分析了偏振分光棱镜、法拉第旋光器以及近似计算这3个主要的测量误差源,并估计总测量误差约为1°。