应用红外光谱显微成像技术观测单细胞雨生红球藻虾青素代谢过程

雨生红球藻是一种适于生产虾青素的单细胞绿藻,虽然已有研究报道了藻细胞内类胡萝卜素的合成代谢途径,但是对类胡萝卜素及虾青素的具体形成过程以及它们在显微尺度上的含量和分布变化还缺乏直接的观测数据和证据。本研究运用高空间分辨率的傅里叶变换红外光谱显微成像技术,对不同时期的单一雨生红球藻细胞中的类胡萝卜素的空间分布和含量进行了观测,根据红外光谱成像数据证实了β胡萝卜素为虾青素合成的前体,并且分析和探讨了细胞中的脂质和蛋白质与虾青素合成及积累的关系。     

应用振动光谱技术检测鉴定微生物的研究进展

振动光谱（红外光谱和拉曼光谱）技术与化学计量学相结合的方法对微生物进行分类、鉴定和无损检测,该方法快速简便、准确度高、仅需微量样品和少量化学试剂、对样品本身没有损害。介绍了振动光谱技术在微生物鉴定检测中的工作原理、关键技术和应用,并对该方法存在的问题和研究前景进行了分析和展望。     

红外光谱图像技术及其在生物学研究中的应用

文章介绍了红外光谱图像技术;介绍了用焦平面阵列检测器、步进扫描傅里叶变换光谱仪、红外显微镜、分束器构成的红外光谱图像硬件系统及信息提取的软件方法;给出了用红外光谱图像分析猴脑组织中蛋白质和磷左异、残留在人胸部组织中的硅酮、水稻叶片等３个在生物学研究中的应用实例;指出了经外光谱图像技术进一步的发展方向是采用同步辐射作为光源、依靠数学算法提高分辨率和发展空间化学计量学来提取空间信息。     

小球藻生长过程脂肪含量动态变化快速无损检测方法研究

微藻-生物柴油转化生产要求产油微藻细胞内大量积累脂肪,而藻类脂肪的积累受外界环境影响较大,因此,微藻生长过程中对藻体脂肪变化进行快速检测和分析有着非常重要的意义。以小球藻(Chlorella sp.)为研究对象,利用可见/近红外光谱技术和高光谱成像技术对不同光源培养条件下微藻生长过程脂肪动态变化和脂肪含量分布可视化分析进行了研究。研究结果表明,虽然利用两种技术获取的小球藻透射光谱和反射光谱有差异,利用连续投影算法进行特征波长也不完全相同,但基于两种技术获得的对于脂肪含量的特征波段光谱建立的小球藻脂肪含量多元线性回归模型的预测结果接近,分别为r_(pre)=0.940,RMSEP=0.003 56和r_(pre)=0.932,RMSEP=0.004 23。研究中小球藻接种初期至生长指数期初期藻体内脂肪含量相对平稳,积累增加发生在生长对数期的末期,而在生长平稳期时的小球藻藻液中,脂肪含量较高的藻体呈现出聚集生长的状态。小球藻生长过程中生命信息快速无损检测方法的实现为微藻实际生产培养和收获策略的制定提供了理论依据和技术手段。     

小球藻、球等鞭金藻和螺旋藻生物量高光谱成像的可视化研究

微藻高效培养是微藻生物能源开发利用的关键和前提,而在营养充足的培养条件下生长迅速但较易受到环境污染和影响,因此微藻生长过程中对其生长状况进行监测意义重大。高光谱成像技术同时拥有丰富物质品质信号的优点和图像包含丰富品质分布空间信息的优点,可为微藻的快速无损检测提供新的方法和手段。分别采集小球藻、球等鞭金藻和螺旋藻三种微藻各45个样本的高光谱图像,并提取样本感兴趣区域(ROI)的平均光谱。利用连续投影算法(SPA)波长优选之后,取30个建模集样本的光谱数据与其相应的生物量建立多元线性回归(MLR)模型,对15个预测集样本的生物量进行预测,小球藻、球等鞭金藻和螺旋藻预测相关系数(r)分别为0.950,0.969和0.961,预测均方根误差(RMSEP)为0.010 2,0.010 7和0.017 1,获得了较好的预测精度。最后,用所建MLR模型对预测集图像上每个像素点的生物量加以预测,采用Matlab图像编程处理将不同的生物量用不同的颜色表示,最终以伪彩图的形式实现藻液生物量的可视化。研究结果表明,高光谱成像技术对小球藻和螺旋藻藻液生物量的可视化效果较好,对球等鞭金藻的预测效果还需要进一步改进。本研究为实现微藻生长信息的快速获取和进一步开展微藻生物质能源利用奠定了一定的研究基础。     

基于高光谱成像技术的斜生四链藻（T. obliquus）碳水化合物和蛋白质判别研究

微藻工厂化养殖为天然碳水化合物、蛋白质等生产提供了重要途径，但较高的养殖成本始终是限制微藻大规模商业化发展的瓶颈之一。由于微藻生长速度很快且其胞内代谢信息时刻都在发生变化，开发快速无损的微藻生长代谢监测手段用以实时获取微藻生产过程中感兴趣指标变化信息，可以据此及时调整培养条件，保障微藻高效优质生产。关于微藻生长代谢信息快速无损检测的研究多集中在微藻油脂及其特性、色素等方面，对于同作为微藻重要营养成分的碳水化合物、蛋白质等却少有报道。本研究以斜生四链藻(Tetradesmus obliquus)为研究对象，利用可见/近红外高光谱成像(HSI)技术结合化学计量学方法，提出基于HSI的微藻碳水化合物和蛋白质反演判别方法。对比研究标准化(autosacling)、标准正态化(SNV)等12种预处理方法对原始高光谱数据的处理效果；采用竞争自适应重加权采样算法(CARS)、区间随机蛙跳算法(iRF)和模拟退火算法(SA)进行特征波段选择；结合多元线性回归(MLR)、偏最小二乘(PLS)、支持向量机回归(SVR)以及随机森林回归(RFR)对微藻生物量、碳水化合物和蛋白质含量进行反演判别。结果表明，斜...     

波谱分析技术在宝石鉴定中的应用评述

波谱分析技术已被成功地应用于宝石学研究和宝石鉴定领域,如红外光谱技术,拉曼光谱技术和顺磁共振技术均可以有效,快速,无损和准确地鉴定宝石的类别,如天然宝石,人工合成宝石和优化处理宝石。     

含油岩心显微红外光谱成像方法的研究

虽然用红外光谱显微探针方法研究岩心中碳质物已有很多工作,但是通常对诸如岩心等样品物性的光谱学微探针实验研究多是局限于对经过复杂处理分离出的微小样品或样品中个别位点所得的结果,缺乏对复杂样品各组分(或基团)的空间分布及其相互关系的研究。近年来新发展起来的光谱成像分析系统将光谱技术与成像技术有机地结合起来融为一体,可在光谱和空间两个方面对目标样品进行分析和识别并已得到广泛应用,但是用红外显微成像光谱研究岩心的工作则少有报道。文章报道了应用透射和衰减全反射(ATR)两种方式对含油岩心进行了显微红外光谱成像(Mapping)的研究。从灵敏度、信噪比、光谱分辨率和空间分辨率以及工作效率等方面对两种方式所得实验结果进行了比较和评估。     

地面长波红外高光谱成像气体探测研究

报道了地面长波红外遥测的新进展 ,具体阐述了窗扫时空调制傅里叶光谱成像技术的实现过程.演示装置基于角锥反射镜M ichelson干涉具 ,构成了空间调制干涉 ;采用了制冷型长波红外焦平面探测器组件 ,通过对数据立方体的采集、重组、基线校正、切趾、相位校正和傅里叶变换等处理 ,实现了长波红外波段高光谱成像.自研的CHIPED-1长波红外高光谱成像原理实验装置的探测灵敏度指标噪声等效辐射通量密度NESR在单次采样时达到了5.6 × 10-8 W · (cm-1 · sr · cm2 )-1 ,与商品化时间调制干涉高光谱成像仪相当 ;反映了技术的先进性 ,并留有较大的改进空间.通过测试聚丙烯薄膜的透过率曲线 ,CHIPED-1红外高光谱成像原理实验装置的光谱响应范围达到了11. 5 μm.文章还以室外高楼和乙醚气体的探测实验为例 ,研究了二维分布化学气体VOC的高光谱成像探测方法.在复杂背景和低试验浓度情况下 ,从同一波数的红外光谱切片上 ,观察不出乙醚蒸气的存在 ,但是进行了差谱处理后 ,可以清楚看到乙醚蒸气的空间分布.高光谱方法应用在有机蒸气VOC的红外探测领域 ,相对于宽波段热成像方法 ,具有灵敏度高、抗干扰能力强和识别种类多等诸多优势.     

小鼠小梁细胞与肌成纤维细胞的同步辐射红外显微光谱

原发性开角型青光眼是常见的致盲性眼部疾病。眼压升高是原发性开角型青光眼发生和发展最主要的危险因素,是由小梁网途径的房水外流排出系统发生病变、房水流出阻力增加所致。研究表明,房水中存在的转化生长因子-β能够使小梁细胞纤维化,诱导小梁细胞过度增殖,从而阻碍房水外流,导致原发性开角型青光眼的发生。原发性开角型青光眼发病隐蔽,病程进展缓慢,早期没有任何症状,往往到晚期视力视野有显著损害时,才会被发现,因此原发性开角型青光眼的早期诊断尤为重要。同步辐射红外显微成像结合高亮度、高分辨率的同步辐射源,同时配备傅里叶变换红外光谱仪与红外显微镜,可以实现细胞的检测。这对从分子层面获取细胞的变化信息,深入理解疾病的发病机制以及疾病的早期诊断具有非常重要的意义。虽然有很多红外光谱在生物医学领域的研究报道,但是应用红外光谱显微成像技术研究细胞等生物医学体系仍然是亟待发展的领域,并且目前未找到关于红外光谱用于小梁网细胞的检测报道。在体外用转化生长因子-β对老鼠小梁网细胞进行诱导,使其转化为肌成纤维细胞,模拟小梁细胞纤维化过程。对小梁网细胞以及经转化生长因子-β诱导形成的肌成纤维细胞进行同步辐射红外显微成像及光谱分析,并进一步探讨同步辐射用于早期诊断原发性开角型青光眼的可行性。研究表明肌成纤维细胞内的弹性蛋白明显高于小梁网细胞,而弹性蛋白中95%为非极性氨基酸,即氨基酸的侧链基团R基只有C和H两种元素。对比两种细胞的红外谱图,发现在2 934,2 900和2 845 cm-1,肌成纤维细胞的 CH₃,CH₂和CH的伸缩振动明显强于小梁网细胞,推测可能是由于转化生长因子-β诱导后细胞内弹性蛋白增加所致。在细胞层面检测了小梁网细胞的过度增殖,为将来可以直接获取细胞的红外光谱从而检测小梁网细胞的增殖程度,进而检测原发性开角型青光眼等疾病奠定了基础。得出同步辐射红外谱学与显微成像有望成为检测原发性开角型青光眼新手段的结论,也为将来便携式红外显微光谱仪临床实时检测青光眼等疾病提供了依据。     

Application of Advanced Synchrotron Radiation–Based and Conventional Molecular Techniques in Recent Research on Molecular Structure,Metabolic Characteristics,and Nutrition in Coproducts from Biofuel Processing

Abstract

Advanced synchrotron radiation infrared microspectroscopy, as a nondestructive and rapid analytical technique, is able to simultaneously reveal the structural, chemical, and environmental features of biomaterials at cellular and molecular levels within intact tissue. However, to date, this advanced synchrotron-based technique is still seldom used by feed and nutrition scientists. This article aims to provide detailed information regarding how to apply advanced synchrotron radiation–based and conventional molecular techniques to research in coproducts from biofuel processing on the molecular structure, metabolic characteristics, and nutrition. The information described in this article provides better insight on coproduct research progress and updates with advanced synchrotron radiation-based and globar-based (conventional) molecular spectroscopy.     

Broad band infrared near-field spectroscopy at finger print region using SPring-8

The authors report infrared near-field spectroscopy using synchrotron radiation at BL43IR, SPring-8 in the finger print region. At the microspectroscopy station, the infrared synchrotron radiation beam is focused on a cantilever probe with a 3 μm square aperture. A comb-shaped Au electrode with the width of 3 μm and the distance of 3 μm is used for the reflection measurement. The Au electrodes can be resolved at 650 cm⁻¹ and the resolution is estimated to be λ/5.     

Performance of the infrared microspectroscopy station at SSRF

At the third generation synchrotron light source Shanghai Synchrotron Radiation Facility (SSRF), the first infrared beamline BL01B has been successfully constructed. The infrared beamline collects both bending magnet and edge radiation. A brief introduction of the infrared beamline design has been given in this article. The infrared microspectroscopy station is equipped with a Nicolet 6700 FTIR spectrometer and a Nicolet Continuum Microscope. The flux at the entrance of the FTIR spectrometer, the intensity profile, the signal to noise ratio (SNR) with different apertures, and the focused spot size of the infrared microspectroscopy station have been measured. The performances with synchrotron radiation infrared source and internal globar source have been compared. The results indicate that the infrared microspectroscopy station at SSRF has the ability of analysis samples in a small area with diffraction limited spatial resolution.     

近红外光谱法对甲醇柴油中甲醇含量测定

应用近红外光谱结合化学计量学方法实现了对甲醇柴油中的甲醇含量的定量分析。以实验室配制的32种不同浓度[浓度范围为2%～25.8%(φ)]的甲醇柴油溶液为研究对象,在4 500～7 000 cm-1光谱范围内,建立偏最小二乘(PLS)、支持向量机(SVM)和最小二乘支持向量机(LS-SVM)三种定量分析模型。在建立SVM模型时,经过比较分析,径向基函数(radial basis function,RBF)作为SVM模型的核函数时可以获得更高的预测精度。最终获得甲醇含量的PLS, SVM和LS-SVM三种模型的预测相关系数R_P分别为0.985 9, 0.990 3, 0.998 9,预测均方根误差RMSEP分别为0.405 2, 0.356 3, 0.062 4,可以看出,三种预测模型都可以达到很好的效果,最优的预测模型是使用LS-SVM建模。研究结果表明,利用近红外光谱法结合化学计量学方法对甲醇柴油中甲醇含量的检测具有可行性,并可以达到很好的效果。采用近红外光谱技术结合化学计量方法对甲醇柴油中甲醇含量进行定量分析,也为近红外光谱技术快速无损检测甲醇柴油甲醇含量提供参考和应用价值。     

Application of Infrared Photoacoustic Spectroscopy in Soil Analysis

Abstract

Soil analysis has become routine work for soil management and crop production. However, laboratory analysis–based determination of soil properties is expensive and time consuming, which is not suitable for precision agriculture. Infrared spectroscopy (IR) appears as an alternative and fast technique to measure soil properties and has had wide application; in particular, a new method called infrared photoacoustic spectroscopy (FTIR-PAS) has been applied in soil analysis. The soil infrared photoacoustic spectrum is more convenient to record; the spectra contain more useful information versus conventional reflectance spectroscopy, and it appears promising for identification of soil types and measure soil properties. The step-scan function of FTIR-PAS makes it possible to explore the soil microstructure in situ; furthermore, more sensible photoacoustic cells (PA), such as a quartz-enhanced PA cell, will make FTIR-PAS a strong tool for the study of soil science. The application of infrared photoacoutic spectroscopy in soil analysis is largely dependent on spectra pretreatment and chemometrics methods due to strong interferences, and more mathematical tools models will benefit or optimize the prediction performance. To make full use of soil infrared spectra, soil spectra library construction is required in the future, which will play an important role in the application of soil analysis.     

Development of scattering near-field optical microspectroscopy apparatus using an infrared synchrotron radiation source

We report the status of a scattering near-field microspectroscopy apparatus developed at SPring-8 using an infrared synchrotron radiation (IR-SR) source. It consists of a scattering type scanning near-field optical microscope and a Fourier transform infrared spectrometer. The IR-SR is used as a highly brilliant and broad-band IR source. This apparatus has potential for application in near-field spectroscopy with high spatial resolution beyond the diffraction limit. In order to eliminate background scatterings from the probe shaft and/or sample surface, we used higher harmonic demodulation method. The near-field spectra were observed by 2nd harmonic components using the lock-in detection. The spatial resolution of about 300 nm was achieved at around 1000 cm^? 1 (10 μm wavelength).     

化学计量学在近红外光谱定性分析中的应用

近红外光谱的多元特性使得它特别适合于应用化学计量学技术进行物质的分析。本文讨论了判别分析技术在近红外光谱定性分析中的应用,结果表明应用相关的化学计量学方法采用近红外光谱进行物质的定性分析是可行的。     

Sensitivity Advantage of QCL Tunable-Laser Mid-Infrared Spectroscopy Over FTIR Spectroscopy

Abstract

Interest in mid-infrared spectroscopy instrumentation beyond classical FTIR using a thermal light source has increased dramatically in recent years. Synchrotron, supercontinuum, and external-cavity quantum cascade laser light sources are emerging as viable alternatives to the traditional thermal black-body emitter (Globar), especially for remote interrogation of samples (“stand-off” detection) and for hyperspectral imaging at diffraction-limited spatial resolution (“microspectroscopy”). It is thus timely to rigorously consider the relative merits of these different light sources for such applications. We study the theoretical maximum achievable signal-to-noise ratio (SNR) of FTIR using synchrotron or supercontinuum light vs. that of a tunable quantum cascade laser, by reinterpreting an important result that is well known in near-infrared optical coherence tomography imaging. We rigorously show that mid-infrared spectra can be acquired up to 1000 times faster—using the same detected light intensity, the same detector noise level, and without loss of SNR—using the tunable quantum cascade laser as compared with the FTIR approach using synchrotron or supercontinuum light. We experimentally demonstrate the effect using a novel, rapidly tunable quantum cascade laser that acquires spectra at rates of up to 400 per second. We also estimate the maximum potential spectral acquisition rate of our prototype system to be 100,000 per second.     

A new beamline for infrared microscopy in the SR center of Ritsumeikan University

A new beamline for an infrared (IR) microspectroscopy is under construction in the SR center of Ritsumeikan University. We designed an optical system which collects synchrotron radiation (SR) photons by installing one toroidal mirror and two plane mirrors in the storage ring chamber. As a result, the acceptance angle can be widened up to 250 mrad in horizontal and 63 mrad in vertical. Our aim is to develop an IR microspectroscopy beamline with spatial resolution of the order of sub micron from mid-IR to far-IR. In this paper, we present a designed optical system of IR microspectroscopy and the results of ray-tracing.