基于NIR-SERS技术结合统计分析肺癌氧合血红蛋白

基于新型、高效、生物兼容性近红外表面增强拉曼散射(NIR-SERS)基底,采用共焦光路设计的便携式拉曼光谱仪,对25例肺癌患者和25例健康人的氧合血红蛋白进行NIR-SERS光谱检测,再对两者NIR-SERS光谱的谱峰强度和谱峰位置进行分析研究。发现两者的NIR-SERS光谱的谱峰强度在一定区域存在较明显的差异性,在473,816和1 583 cm-1处健康人的光谱的谱峰强度比肺癌患者要大,健康人氧合血红蛋白平均NIR-SERS光谱在1 208和1 425 cm-1谱峰处时,肺癌患者分别蓝移到1 212和1 432 cm-1,健康人氧合血红蛋白平均NIR-SERS光谱在1 583 cm-1的谱峰处,肺癌患者红移到1 579 cm-1。采用主成分(PCA)分析和独立变量T检验统计分析方法得出NIR-SERS光谱在主成分得分PC3,PC4和PC9处具有显著差异,并通过PCA得分二维、三维散点图可以将健康人和肺癌患者区分,诊断的特异性为88%,灵敏度为80%,总判别正确率为84%。通过谱峰归属分析,两者的NIR-SERS光谱存在显著差异与氧合血红蛋白吡咯环和乙烯基团(C_aC_m)的振动有关。该研究为肺癌的SERS光谱技术诊断提供了实验依据,NIR-SERS技术有望发展成为一种新型的临床辅助诊断工具。     

基于静电自组装银膜的急性粒白血病氧合血红蛋白表面增强拉曼光谱研究

文中基于高效、生物兼容性纳米结构银膜,采用便携式拉曼光谱仪分别对10个健康人和10个急性粒白血病患者的氧合血红蛋白进行了表面增强拉曼散射(SERS)光谱的研究。实验发现,健康人与急性粒白血病患者的氧合血红蛋白SERS光谱存在显著差异:(1)健康人氧合血红蛋白SERS光谱中位于340cm-1附近很弱的拉曼峰在急性粒白血病患者的SERS光谱中变的很强;(2)健康人氧合血红蛋白SERS光谱中位于655 cm-1附近的拉曼峰在急性粒白血病患者SERS光谱中蓝移到670 cm-1附近,同时在727cm-1附近出现一个新的拉曼峰;(3)对健康人和急性粒白血病患者氧合血红蛋白SERS光谱中位于472、814、1335、1423和1588 cm-1处的拉曼峰相对强度比研究发现,相对强度比I814/I472,I1335/I472、I1423/I472和I1588/I472可以作为较好的区分健康人和急性粒白血病患者的SERS光谱诊断指标,这为基于SERS光谱技术进行急性粒白血病诊断提供了初步实验依据。     

基于NIR-SERS技术结合PCA-LDA统计分析肝癌术后恢复状况

采用近红外表面增强拉曼散射(NIR-SERS)光谱技术,基于高效、稳定、低廉的纳米银膜基底检测了25例肝癌患者、25例肝癌术后患者和25例健康人的氧合血红蛋白(Oxy Hb)。对比发现,健康人、肝癌术后患者和肝癌患者的Oxy Hb的NIR-SERS光谱谱峰强度呈现依次下降的趋势,表明从谱峰强度可以初步判断肝癌术后患者恢复情况。利用主成分分析(PCA)结合线性判别分析(LDA)统计方法进行分析,25个肝癌术后患者中8人判别为健康人,其余17人判别为肝癌患者。判别分组与实际情况相符,表明PCA-LDA统计方法可以更准确地判断肝癌术后患者恢复状况。因此,NIR-SERS技术结合PCA-LDA统计方法有望为肝癌术后患者恢复状况的诊断提供一种新的思路和方法。     

胃癌患者血红蛋白的表面增强拉曼光谱分析

胃癌患者能够得到早期诊断对其治疗具有十分重要的意义,疾病状态下的血红蛋白拉曼光谱检测在血液代用品的高铁血红蛋白含量检测中以及血氧饱和定量测定中占很大优势。本实验采用微波加热法制备银胶体粒子,依次对20例胃癌患者和11例健康人的血红蛋白进行表面增强拉曼光谱分析。采用SERS谱峰归属分析结果显示胃癌患者血红蛋白中的酪氨酸、苯丙氨酸和吡咯环的含量均低于正常人。本文还讨论了血红素的分子结构,在血红蛋白和氧气结合的前后,Fe2+分别处于高低自旋态,离子半径也随着与氧结合缩小了0.075 nm从而滑落入卟啉环平面中央的孔隙之中。这种空间的拉伸变化会牵动与铁相连的F8His,使得珠蛋白中两股螺旋之间空隙缩小,导致将HC2酪氨酸排挤出空隙。利用这一机理,对血红蛋白1 560 cm-1的吸收峰进行研究,证实了胃癌患者酪氨酸含量确实低于正常人。为了能得到更加明显的诊断区分,利用降维的思想,采用主成分分析(PCA)的方法对所有的拉曼光谱进行多元统计分析,得出三维的诊断散点图。为了更加精确的得出诊断准确率,用判别分析得出诊断的灵敏度和特异性分别为90.0%和90.9%,总判别正确率为90.3%。此项研究表明：对氧合血红蛋白的表面增强拉曼光谱诊断分析有希望成为一项新型的胃癌诊断技术应用于医学领域。     

鼻咽癌患者唾液的表面增强拉曼光谱研究

表面增强拉曼光谱(SERS)技术是一种快速、有效、无损的光学诊断方法,能提供生物分子成分和结构的指纹特征。人的唾液包含多种能反应人体健康状况的生物标志物。采用纳米银溶胶为增强基底的SERS技术,从46位鼻咽癌患者、30位鼻炎患者和48位健康人的唾液样本中采集了208条高质量的SERS。研究结果表明鼻咽癌患者、鼻炎患者和健康人的唾液SERS之间存在明显差异。利用主成分分析结合线性判别分析(PCA-LDA)对采集的唾液SERS进行诊断研究,获得灵敏度分别为89.5%,93.0%和100%,特异性分别为94.6%,96.7%和97.7%。研究结果表明：唾液SERS技术结合PCA-LDA分析方法可能快速、有效区分鼻咽癌、鼻炎患者与正常人的唾液样本,具有鼻咽癌无损检测和临床筛查的潜能。     

应用血红蛋白的拉曼光谱进行食管癌诊断的研究

早期诊断对食管癌患者的治疗具有重要意义。通过对21例食管癌患者和20例健康人的血红蛋白溶液进行拉曼光谱的检测和研究,发现食管癌患者与健康人血红蛋白的拉曼光谱有较明显的区别。与健康人相比,食管癌患者血红蛋白中处于低自旋态的铁离子增多,而处于高自旋态的铁离子减少,说明癌变导致食管癌患者血红蛋白中部分处于高自旋态的铁离子转移到了低自旋态,这与癌症患者血样较易溶血的现象相吻合。采用主成分分析(PCA)方法,对所得拉曼光谱建立多元统计模型进行计算,得到的光谱峰值的PCA得分三维散点图可以良好的区分食管癌患者与健康人。利用判别分析方法得出该算法诊断食管癌的灵敏度和特异性分别为90.5%和95%,总判别正确率为92.7%。结果表明,对血红蛋白的拉曼光谱进行检测和分析有可能成为食管癌早期诊断的有效工具之一。     

基于SERS的一种快速、有效的挥发物筛选式葱属植物鉴别分类研究

本文应用表面增强拉曼散射技术以纳米银溶胶作为基底直接对17种葱属植物的挥发性物质进行了检测,进一步用SERS谱图结合化学计量学多变量统计分析,对17种葱属植物进行鉴别分类研究,并提出一种基于SERS的快速、有效的挥发性物质筛选式葱属植物鉴别分类研究方法。对不同年份制作的纳米银溶胶进行了重现性测试,结果显示纳米银溶胶作为SERS基底对葱属植物的挥发性物质检测重现效果较好;对同一植物不同部位的挥发性物质进行检测,结果显示光谱峰位变化不大,只是个别峰的相对强度发生了变化;对17种葱属植物的挥发性物质进行检测,结果显示:17种葱属植物的挥发物的SERS光谱可分为三组,1-丙硫醇增强组、烯丙基甲基硫醚增强组、二烯丙基二硫增强组,说明纳米银溶胶对葱属植物的挥发物具有选择性增强效果;17种葱属植物挥发物的SERS谱结合聚类分析、因子分析、判别分析进行多变量统计分析,分析结果显示,样品能按三个不同增强组进行准确分类。实验结果表明,基于SERS的挥发物筛选式葱属植物鉴别分类研究方法可以为葱属植物分类研究提供参考信息。     

基于新型NIR-SERS基底的肝癌血清NIR-SERS光谱研究

基于一种新型、高效、生物兼容性近红外表面增强拉曼散射(NIR-SERS)基底,采用便携式近红外拉曼光谱仪分别对健康人和肝癌病患者的血清进行了NIR-SERS光谱研究.实验发现,健康人与肝癌患者的血清NIR-SERS光谱存在显著差异:1)健康人血清NIR-SERS光谱中位于630、720、812和1578 cm-1附近的...     

小波变换基线扣除对激光诱导等离子体温度计算的改善

温度是激光诱导等离子体特性研究最重要的参数之一,为降低光谱连续背景对Boltzmann平面法计算等离子体温度的精密度的影响,利用小波变换对等离子体光谱进行分解,并采用软阈值法对代表光谱基线的低频信号进行扣除。选择合适的小波分解层数L及阈值系数α能有效提高Boltzmann图的线性相关度,即有更高的拟合系数R2,从而提高等离子体温度的计算精密度。对低合金钢样品417～445 nm波段LIBS光谱采用db4小波函数进行分解、基线扣除和信号重构,选用12条Fe原子谱线建立Boltzmann图,由Boltzmann图拟合直线的斜率计算得到等离子体温度。对L和α系数进行了优化选择,研究发现,采用8层小波分解时Boltzmann图具有较高的R2,而α的选择与时延t_d有关,t_d≤4.0 μs时,α=0.3可获得最佳R2值,之后随t_d的增大,α逐渐减小; 在t_d≥6.5 μs后,α=0,即光谱低频信号被完全扣除,说明基线对光谱特征谱线的干扰随时延的增加逐渐减弱。基线扣除后各时延的等离子体温度降低约2 000～3 000 K,温度随时延的增加逐渐降低,与等离子体膨胀过程中温度逐渐下降的物理过程相吻合,且变化过程中的波动变小。     

联用多种荧光光谱研究高铁酸钾对荧蒽的降解

应用多种荧光光谱追踪了高铁酸钾对荧蒽的降解过程, 包括发射, 同步, 三维(EEM), 时间扫描及光度定量所提供丰富的关于整个降解反应的信息, 探讨了高铁酸钾对荧蒽分子的降解特性及其随时间变化规律; 分别对该降解过程中的不同时段(20, 40, 70 s)的时间扫描荧光曲线进行拟合,并由此推测了高铁酸钾降解荧蒽的动力学特征。结果显示, 相同的反应时段, 各荧光光谱反映的高铁酸钾对荧蒽分子的降解率非常接近: t=10 s约55%; t=25 s约81%; t=40 s约91%。各时段的降解过程中均未出现新的荧光特性; t≤20 s, 为该降解反应的决定性阶段, 此阶段的降解过程最为接近线性关系; t>20 s, 降解过程偏离线性关系越来越远。整个降解过程可以近似地用一级反应动力学规律描述。     

Near‐infrared surface‐enhanced Raman spectroscopy (NIR‐SERS) studies on oxyheamoglobin (OxyHb) of liver cancer based on PVA‐Ag nanofilm

A near‐infrared surface‐enhanced Raman spectroscopy (NIR‐SERS) method was employed for oxyheamoglobin (OxyHb) detection to develop a simple blood test for liver cancer detection. Polyvinyl alcohol protected silver nanofilm (PVA‐Ag nanofilm) used as the NIR‐SERS active substrate to enhance the Raman scattering signals of OxyHb. High quality NIR‐SERS spectrum from OxyHb adsorbed on PVA‐Ag nanofilm can be obtained within 16 s using a portable Raman spectrometer. NIR‐SERS measurements were performed on OxyHb samples of healthy volunteers (control subjects, n = 30), patients (n = 40) with confirmed liver cancer (stage I, II and III) and the liver cancer patients after surgery (n = 30). Meanwhile, the tentative assignments of the Raman bands in the measured NIR‐SERS spectra were performed, and the results suggested cancer specific changes on molecule level, including a decrease in the relative concentrations and the percentage of aromatic amino acids of OxyHb, changes of the vibration modes of the C_aH_m group and pyrrole ring of OxyHb of liver cancer patients. In this paper, principal component analysis (PCA) combined with independent sample T test analysis of the measured NIR‐SERS spectra separated the spectral features of the two groups into two distinct clusters with the sensitivity of 95.0% and the specificity of 85.7%. Meanwhile, the recovery situations of the liver cancer patients after surgery were also assessed using the method of discriminant analysis‐predicting group membership based on PCA. The results show that 26.7% surgeried liver cancer patients were distinguished as the normal subjects and 63.3% were distinguished into the cancer. Our study demonstrated great potentials for developing NIR‐SERS OxyHb analysis into a novel clinical tool for non‐invasive detection of liver cancers. Copyright © 2013 John Wiley & Sons, Ltd.     

Label‐free optical detection of type II diabetes based on surface‐enhanced Raman spectroscopy and multivariate analysis

Surface‐enhanced Raman scattering (SERS) spectroscopy was first employed to detect oxyhemoglobin (OxyHb, the common type of hemoglobin) variation in type II diabetic development without using exogenous reagents. Using silver nanoparticles as SERS‐active substrate, high‐quality SERS spectra are obtained from blood OxyHb samples of 49 diabetic patients and 40 healthy volunteers. Tentative assignment of the observed SERS bands indicates specific structural changes of OxyHb molecule in diabetes, including heme transformation and globin variation. Furthermore, partial least squares and principal component analysis combined with linear discriminate analysis diagnostic algorithms are employed to analyze and classify the SERS spectra acquired from diabetic and healthy OxyHb, yielding the diagnostic accuracies of 90.0% and 95.5%, respectively. This exploratory work suggests that the silver nanoparticles‐based OxyHb SERS method in combination with multivariate statistical analysis has great potential for the label‐free detection of type II diabetes. Copyright © 2014 John Wiley & Sons, Ltd.     

Different classification algorithms and serum surface enhanced Raman spectroscopy for noninvasive discrimination of gastric diseases

In this study, surface enhanced Raman spectroscopy (SERS) was used to investigate the spectral characteristics of blood serum for the purpose of diagnosing stomach diseases. SERS spectral data was collected from patients with atrophic gastritis, both pre‐operation and post‐operation gastric cancer, and from healthy individuals. Visual differences in the SERS spectra were observed between the four groups which indicate corresponding biomolecule concentration changes in blood. To further investigate the diagnostic ability of human serum, the spectral data was analyzed with three chemometric processes. These three methods extracted features and classified from the spectral data. Principal component analysis (PCA) was first performed to reduce the dimensionality of the original spectral data. Then, the classification methods support vector machine (SVM), linear discriminant analysis (LDA) and classification and regression tree (CART) were used for the evaluation of diagnostic ability. Accuracies of 96.5%, 88.8% and 87.1% were obtained for PCA‐SVM, PCA‐LDA and PCA‐CART, respectively. Copyright © 2016 John Wiley & Sons, Ltd.     

皮疹病人血清的表面增强拉曼光谱

研究了皮疹患者和健康人血清的表面增强拉曼光谱。结果表明： 皮疹患者血清中蛋白质主链酰胺Ⅰ的谱线出现在1 648 cm-1处,而在健康人血清中此峰消失。蛋白质侧链C—S键扭曲构象的谱线637 cm-1的相对强度增加了23%,而反式构象的谱线725 cm-1的相对强度减少了60%,说明C—S键的反式构象部分转成扭曲构象,表明皮疹患者血清中蛋白质的有序结构发生变化。类脂物特征峰1 449 cm-1的相对强度增加了近一倍,D-甘露糖的谱线1 099 cm-1消失,表明患者血清中糖蛋白、糖质发生了变化。这些拉曼特征峰为皮疹的诊断以及生化机理研究提供了有力的实验依据。最后用主成分分析(PCA)方法对健康人和皮疹患者血清的表面增强拉曼光谱进行分析,发现皮疹患者血清的主成分分布相对健康人较分散,并且此方法能准确的区分开这２种血清。     

Surface‐enhanced Raman scattering spectroscopy for potential noninvasive nasopharyngeal cancer detection

Combining membrane electrophoresis with surface‐enhanced Raman scattering (SERS) spectroscopy, the serum proteins were first purified and then mixed with silver nanoparticles to perform SERS spectral analysis. Therefore, the spectral signatures were enhanced to high‐fidelity SERS signatures because of the purification procedure of the first step. We used the method to analyze blood plasma samples from nasopharyngeal cancer patients (n = 43) and healthy volunteers (n = 33) for cancer detection. Principle component analysis of the SERS spectra revealed that the data points for the cancer group and the normal group form distinct, completely separated clusters with no overlap. Therefore, the nasopharyngeal cancer group can be unambiguously discriminated from the normal group, i.e., with both diagnostic sensitivity and specificity of 100%. These results are very promising for developing a label‐free, noninvasive, and reliable clinical tool for rapid cancer detection and screening. Copyright © 2011 John Wiley & Sons, Ltd.