Experimental and statistical analysis methods for peptide detection using surface‐enhanced Raman spectroscopy

Surface‐enhanced Raman spectroscopy (SERS) has the potential to make a significant impact in biology research due to its ability to provide information orthogonal to that obtained by traditional techniques such as mass spectrometry (MS). While SERS has been well studied for its use in chemical applications, detailed investigations with biological molecules are less common. In addition, a clear understanding of how methodology and molecular characteristics impact the intensity, the number of peaks, and the signal‐to‐noise of SERS spectra is largely missing. By varying the concentration and order of addition of the SERS‐enhancer salt (LiCl) with colloidal silver, we were able to evaluate the impact of these variables on peptide spectra using a quantitative measure of spectra quality based on the number of peaks and peak intensity. The LiCl concentration and order of addition that produced the best SERS spectra were applied to a panel of synthetic peptides with a range of charges and isoelectric points (pIs) where the pI was directly correlated with higher spectral quality. Those peptides with moderate to high pIs and spectra quality scores were differentiated from each other using the improved method and a hierarchical clustering algorithm. In addition, the same method and algorithm was applied to a set of highly similar phosphorylated peptides, and it was possible to successfully classify the majority of peptides on the basis of species‐specific peak differences. Copyright © 2008 John Wiley & Sons, Ltd.     

Effect of an aliphatic spacer group on the adsorption mechanism on the colloidal silver surface of L‐proline phosphonodipeptides

A comparative study of molecular structures of five L ‐proline (L ‐Pro) phosphonodipeptides: L ‐Pro‐NH‐C(Me,Me)‐PO₃H₂ (P1), L ‐Pro‐NH‐C(Me,iPr)‐PO₃H₂ (P2), L ‐Pro‐L ‐NH‐CH(iBu)‐PO₃H₂ (P3), L ‐Pro‐L ‐NH‐CH(PA)‐PO₃H₂ (P4) and L ‐Pro‐L ‐NH‐CH(BA)‐PO₃H₂ (P5) has been carried out using Raman and absorption infrared techniques of molecular spectroscopy. The interpretation of the obtained spectra has been supported by density functional theory calculations (DFT) at the B3LYP; 6–31 + + G** level using Gaussian 2003 software. The surface‐enhanced Raman scattering (SERS) on Ag‐sol in aqueous solutions of these phosphonopeptides has also been investigated. The surface geometry of these molecules on a silver colloidal surface has been determined by observing the position and relative intensity changes of the Pro ring, amide, phosphonate and so‐called spacer (−R) groups vibrations of the enhanced bands in their SERS spectra. Results show that P4 and P5 adsorb onto the silver as anionic molecules mainly via the amide bond (∼1630, ∼1533, ∼1248, ∼800 and ∼565 cm⁻¹), Pro ring (∼956, ∼907 and ∼876 cm⁻¹) and carboxylate group (∼1395 and ∼909 cm⁻¹). Coadsorption of the imine nitrogen atom and PO group with the silver surface, possibly by formation of a weaker interaction with the metal, is also suggested by the enhancement of the bands at 1158 and 1248 cm⁻¹. P1, P2 and P3 show two orientations of their main chain on the silver surface resulting from different interactions of the  C CH₃,  NH and  CONH fragments with this surface. Bonding to the Ag surface occurs mainly through the imino atom (1166 cm⁻¹) for P2, while for P1 and P3 it occurs via the methyl group(s) (1194–1208 cm⁻¹). The amide group functionality (CONH) is practically not involved in the adsorption process for P1 and P2, whereas the C_s P bonds do assist in the adsorption. Copyright © 2008 John Wiley & Sons, Ltd.     

高层大气风场四强度测量法误差分析与计算

利用干涉成像光谱技术和电磁波的多普勒效应,通过测量高层大气(80—300km)中的气辉(极光)辐射线的四个干涉强度值而获知高层大气的速度和温度信息.采用四强度干涉测量法,分别对基于Michelson干涉仪的动镜扫描和基于无动镜四分区镀膜的干涉图获取模式所引起的测量误差进行了深入的理论分析与研究.给出了由于此测量误差所导致的相位误差所引起的高层大气风场速度、温度的误差数值;采用计算机模拟得出了在不同相位误差条件下,两种模式计算得到的风场速度和温度的误差分布图;给出了风速误差、温度误差与相位步进误差的关系曲线 关键词： 高层大气风场测量 四强度测量法 模拟计算     

硅薄膜沉积过程中等离子发光基团的一维空间分布研究

使用光发射谱(OES)对甚高频等离子增强化学气相沉积(VHF-PECVD)技术沉积硅薄膜时的等离子体发光基团的空间分布进行了在线监测和研究. 研究表明：等离子体的不同发光基团都存在着一个中间强度较大的区域和两边电极附近的暗区;增大硅烷浓度和提高辉光功率都会增大SiH^*峰强度;硼烷的加入,使得SiH^*和Hα^*峰强度增大,但硼烷流量变化的影响很小;硼烷流量增大,材料的晶化率下降,而I_{［Hα^*］关键词： 甚高频等离子增强化学气相沉积 等离子体 发光基团 空间分布}     

大功率单频多芯光纤放大器中抑制受激布里渊散射的分析

针对多芯光纤完善了描述抽运光、信号光和Stokes信号的速率方程组.考虑了温差对受激布里渊散射的影响,利用有限元法求解温度分布方程组,分析了前向和后向抽运方式、对流系数、Stokes初始功率、光纤掺杂粒子密度和光纤长度对受激布里渊散射增益的影响.研究表明：后向抽运方式在抑制受激布里渊散射方面具有明显优势;减小对流系数有助于抑制受激布里渊散射;提高光纤掺杂密度能够加强抑制受激布里渊散射,同时也有助于提高光纤放大器的斜率效率.比较了在相同最佳光纤长度条件下,单芯和19芯光纤放大器的最高工作温度和受激布里渊散射 关键词： 光纤放大器 受激布里渊散射 大功率 有限元法     

受激布里渊散射新介质——全氟胺的研究

本文从介质化学结构与受激布里渊散射(SBS)特性的关系入手,寻找出了SBS特性良好的全氟胺系列新介质——FC-131,FC-3283,FC-40,FC-43,FC-70等,并测定或计算出了新介质的SBS参数. 结果表明,新介质的吸收系数均小于10^-3 cm^-1,光学击穿阈值均高于100 GW/cm². 全氟胺系列新介质不仅具有良好的SBS特性,而且还具有无毒、低挥发性和高稳定性等一系列独特的物理化学性质. 新介质的发现不仅增加SBS介质的种类,而且能够有效提高SBS系统的性能,对于SBS相位共轭镜在高功率激光系统中的应用打下了良好的基础. 关键词： 受激布里渊散射(SBS) 全氟胺 吸收系数 光学击穿阈值     

Adsorption of 2‐amino‐5‐cyanopyridine on a gold surface as probed by surface‐enhanced Raman spectroscopy

The adsorption of 2‐amino‐5‐cyanopyridine (2‐ACP) was investigated in solution at different pH values by i n situ surface‐enhanced Raman scattering (SERS) spectroscopy combined with the electrochemical method. The assignments of the main bands were first performed on the basis of the spectral features of similar compounds and with the help of density functional theory calculations. The results revealed that the adsorption and the interfacial structure of 2‐ACP on the Au electrode depended on the applied potential and the pH values of the solution. In the natural solution, 2‐ACP was adsorbed on the surface with a vertical orientation by the CN group from − 0.4 to − 1.0 V, whereas in the − 0.4 to 0.8 V range, the N atom of the pyridine ring was bound to the surface. A transition region for the reorientation of the two adsorption modes was observed from − 0.8 to − 0.4 V. A flat configuration was preferred at an extremely negative potential. A similar surface adsorption behavior was observed in the alkali environment, while the Stark effect slope decreased because of the adsorption of OH⁻. Due to the protonation of N atom in the acidic solution, the potential region for the coexistence of two configurations ranged from − 0.4 to 0.2 V. Additionally, a similar adsorption configuration was proposed on the Au colloids at various pH values. The results revealed that the adsorption behavior became more complex on colloidal surfaces than that on a rigid electrode surface. Copyright © 2010 John Wiley & Sons, Ltd.     

Enhancing and suppressing four‐wave mixing in electromagnetically induced transparency window

We demonstrate the enhancement and suppression of four‐wave mixing (FWM) in an electromagnetically induced transparency (EIT) window in Y‐type ⁸⁵Rb atomic system. The generated two‐photon FWM signal can be selectively enhanced and suppressed via an EIT window. The EIT of probe as well as the enhancement and suppression of FWM signals can be modified by the sequential‐cascade double dressing. The influence of different probe polarization configurations is also studied. Different polarization states of the probe laser can select different transitions among Zeeman sublevels and different dressing strengths. Copyright © 2010 John Wiley & Sons, Ltd.     

Rapid detection of a foreign protein in milk using IMS–SERS

A method for rapid detection of foreign protein contamination in complex food matrices is critically needed. Here we present a novel method that combines immunomagnetic separation (IMS) and surface‐enhanced Raman scattering (SERS) to detect ovalbumin (OVA), an egg white protein, added into whole milk. IMS was used to specifically capture the OVA out of the milk. Then SERS was applied to analyze the IMS eluate using silver dendrites as the substrate. Two SERS sample preparation methods, namely solution based and substrate based, were used to prepare the IMS eluate for SERS analysis. Results show both methods were able to detect 1 µg OVA in 1 ml milk (1 part per million). Based on the results of principal component analysis and partial least‐squares analysis, solution SERS was more capable of quantitative analysis, while substrate SERS was more sensitive for qualitative analysis. The total analytical time for IMS–SERS was less than 20 min, which satisfied the requirement of rapid detection in a milk processing facility. Copyright © 2011 John Wiley & Sons, Ltd.     

Tip‐heating‐assisted Raman spectroscopy at elevated temperatures

We demonstrate tip‐heating‐assisted enhanced Raman spectroscopy to investigate the temperature dependence of the carbon nanotube G‐band with nanoscale resolution. The controllable and nanoscale heat generated at the tip apex was used to thermally perturb and characterize a small volume in a carbon nanotube sample that is precisely positioned underneath the tip. The dependence of tip enhancement with temperature was also experimentally examined, which is in good agreement with the enhancement calculated from the electromagnetic model of isolated spheroids. The technique presented may open up opportunities in the study of controlled heat‐assisted biochemical reactions and physical transformations of nanostructures. It can also be used for thermal characterization of various materials requiring site‐selective and controllable nanoscale heat source and could enable the realization of new photothermal devices. Copyright © 2010 John Wiley & Sons, Ltd.