高光谱成像技术在生物医学中的应用

高光谱成像技术是传统成像与光谱技术相结合的一门新技术,其可同时获得被测物体的空间特征与光谱信息,以实现对物质特性的研究。本文介绍了高光谱成像技术的基本原理、系统的基本构成及特点,总结和阐述了近年来高光谱成像技术在生物医学领域的发展,以及其在疾病诊断中的应用。     

基于可见/近红外高光谱成像技术的西甜瓜糖度检测

提出了以不同品种的西瓜、甜瓜为研究对象,利用可见光/近红外成像高光谱技术分析不同糖度的西瓜、甜瓜的光谱差异及西瓜、甜瓜糖度在可见光/近红外波段范围的响应。研究表明应用成像高光谱技术检测西甜瓜糖度具有可行性,为进一步研究不同水果糖度高精度模型奠定基础。     

基于高光谱成像和主成分分析的水稻茎叶分割

在单株水稻表型测量研究中,为了实现绿叶面积和茎叶相关表型参数的准确计算提供技术保障,茎叶的分割是非常重要的一步。传统的人工测量方法费时费力,且主观性较强,而基于普通相机拍摄的彩色图像进行分割效果很差。本研究介绍了一种使用可见光-近红外高光谱成像系统自动区分单株盆栽水稻茎叶的方法。首先将各波长下的图像从原始二进制数据中提取出来,接着使用主成分分析所有波长下的图像,并提取出主要的主成分图像,再基于数字图像处理技术将茎叶区分开。实验结果表明,本系统以及文中所用方法对分蘖盛期的水稻茎叶有很好的分割效果,这为后续水稻茎叶表型性状高通量、数字化、无损准确提取提供了重要的技术保障,并进一步促进植物表型组学的发展。     

多光谱成像技术在生物医学中的应用进展

多光谱成像(multispectral imaging,MSI)技术在生物医学可视化方面是一种新技术,它结合了两个已建立的光学模块:成像学和光谱学。它的原理是基于液晶可调谐滤光片,从可见光到近红外波长(400-970nm)区域获取多光谱图像。自从MSI系统加上显微镜商品化以来,MSI已经成为一种快速发展的领域,可应用于细胞生物学、临床前药物开发和临床病理学等。国外已有大量关于MSI在生物医学中应用的研究报道,但国内报道少见。本文主要对多光谱成像的基本原理,近三年内该技术在生物医学领域的应用进展作一简要综述。     

滨海盐土土壤水分的高光谱参数及估测模型

基于滨海盐土5个试验点的土壤含水量和室内土壤表面高光谱反射率,综合分析了350～2500 nm波段范围内土壤含水量与土壤光谱之间的关系,并基于比值光谱指数(RSI)、归一化光谱指数(NDSI)和差值光谱指数(DI)确定了光谱参数,进而构建土壤含水量估测定量模型.结果表明： 滨海盐土原始光谱反射率与土壤含水量呈显著负相关关系,且最大负相关出现在1930 nm(r=0.86)附近.对RSI、NDSI和DI的直线回归方程、幂函数回归方程进行对比,以RSI(R₁₄₀₇,R₁₄₅₉)为自变量构建的土壤含水量指数函数线性回归方程决定系数最大(0.780),标准误较小(0.016),拟合方程为y=0.00001e^9.72053x.估测模型能够更好地监测滨海盐土土壤水分状况.基于RSI(R₁₄₀₇,R₁₄₅₉)构建的模型可实现对江苏省滨海盐土土壤水分的精确监测.     

多光谱成像技术在植物学研究中的应用

多光谱成像(MSI)技术是一种新兴的成像检测技术, 通过将光谱与成像合二为一, 可实现植物结构、生理、生化表型的定性定量分析及其特征分布的评估。近年来, 与数学建模分析结合的MSI技术具有强大的信息捕获能力, 在植物学研究中展现出强劲的应用潜力。该文介绍了MSI技术的成像原理, 总结了近年来MSI技术在植物损伤鉴定、病害研究、代谢物质生化特征及生理进程鉴定方面的应用, 展望了该技术在植物研究领域的前沿性发展, 以期使MSI技术在植物研究中得到更好的应用。     

烟蚜为害特征的高光谱比较

利用手持式高光谱仪测定不同蚜量为害后烟草冠层光谱反射率的变化,同时使用叶绿素计测量叶绿素含量。结果表明,烟草光谱呈现标准植被光谱曲线,烟蚜Myzuspersicae(Sulzer)为害造成烟草光谱反射率下降,近红外波段反射率下降更为明显。和健康烟株相比,轻度(单株顶尖和上部5片叶蚜量≤15头)、中度(15头<单株顶尖和上部5片叶蚜量≤50头)、重度(单株顶尖和上部5片叶蚜量>50头)危害在绿光波段反射率分别下降12%,27%,52%,在近红外波段分别下降15%,20%,38%。对反射率曲线进行微分分析,健康烟株、轻度、中度和重度为害后的一阶导数最大值分别为0.031,0.022,0.026和0.019,其值随着蚜量增加而下降,对应的波长重度为害为718.45nm,其它为716.91nm,红边位置向长波方向移动1.46nm。蓝光、绿光、红光和近红外4个波段反射率和叶绿素含量SPAD值存在良好的线性回归关系,相关系数都达到极显著,绿光波段反射率对叶绿素含量的回归关系尤为显著;同时SPAD值越大,光谱反射率越高。该研究对于应用遥感技术大面积监测烟草生产中病虫害的发生,确定防治适期以及适宜的防治措施具有重要的应用前景。     

基于光谱编码的血管内光声组分成像

光声成像突破了传统的光学成像和超声成像在生物组织成像领域的困境,该技术基于光声(Photoacoustic,PA)效应,脉冲激光激励下的生物组织产生超声信号,超声信号被接收后,通过反投影算法将其携带的时间信息和强度信息转化为能够反映生物组织吸收结构和分布的可视化图像。基于不同生物组织的光吸收差异,当激发光强度均匀且稳定时,光声成像反映的就是该物质对于该波长光的吸收特性。本文中,我们基于导管式的血管内光声断层扫描平台结合多波长激发的光声成像算法开发了基于光谱编码的血管内光声组分成像系统,实现了在离体血管斑块中脂质组分的定量成像,高分辨获得了脂质核心的大小形态和边界信息,表征了斑块内的脂质相对含量。     

基于因子分析的苜蓿叶片叶绿素高光谱反演研究

因子分析是一种能够将具有错综复杂关系的变量归结为少数几个综合因子的多变量统计分析方法,在降低数据维数的同时又可以保存足够的信息,这为处理信息量丰富但冗余较大的高光谱数据提供了一种有效方法。本文利用2010年9月23日采集的16个样点的苜蓿叶片反射率及叶绿素含量数据,采用因子分析方法,分别提取苜蓿叶片反射率光谱400～900nm,以及可见光400nm～760nm和近红外760～900nm光谱区的公共因子,分析因子载荷分布、载荷总量对公共因子与叶绿素含量相关性的影响。利用逐步回归法建立基于公共因子的叶片叶绿素反演模型,并将反演模型与光谱指数建立的模型进行对比。研究表明,1）公共因子与叶片叶绿素的相关性,在更大程度上是与该因子在各个波段上载荷分布有关,而不是总载荷量;2）对波谱进行分区建立的反演模型略优于全区因子分析建立的反演模型;3）与常用于叶片叶绿素含量反演的光谱指数CARI、MCARI、mND680、mND705、mSR705、TVI、DmSR、BGI、BRI相比,因子分析建立的叶绿素反演模型精度更高。     

橡胶树叶片高光谱特征分析

从光谱曲线特征和光谱变换特征分析橡胶树(Hevea brasiliensis)叶片反射曲线特征.结果表明,蓝边、红边、黄边位置特征分别出现于525 nm、725 nm、550 nm波段附近,红谷位置特征变化较大,并提取了红边积分面积等重要光谱变量特征.叶片氮含量与反射光谱的相关分析表明橡胶树叶片氮素敏感波段为700～1300 nm,其中730 nm处相关性最好,达到0.8422的极显著水平,以730 nm处的反射率与叶片氮含量建立线性模型,其复相关系数R2达到0.7094.     

Investigation of human pancreatic cancer tissues by Fourier Transform Infrared Hyperspectral Imaging

Fourier‐transform infrared hyperspectral imaging (FTIR‐HSI) provides hyperspectral images containing both morphological and chemical information. It is widely applied in the biomedical field to detect tumor lesions, even at the early stage, by identifying specific spectral biomarkers. Pancreatic neoplasms present different prognoses and are not always easily classified by conventional analyses. In this study, tissue samples with diagnosis of pancreatic ductal adenocarcinoma and pancreatic neuroendocrine tumor were analyzed by FTIR‐HSI and the spectral data compared with those from healthy and dysplastic samples. Multivariate/univariate approaches were complemented to hyperspectral images, and definite spectral markers of the different lesions identified. The malignant lesions were recognizable both from healthy/dysplastic pancreatic tissues (high values of phospholipids and triglycerides with shorter, more branched and less unsaturated alkyl chains) and between each other (different amounts of total lipids, phosphates and carbohydrates). These findings highlight different metabolic pathways characterizing the different samples, well detectable by FTIR‐HSI.     

Differentiation of Wheat Diseases and Pests Based on Hyperspectral Imaging Technology with a Few Specific Bands

Hyperspectral imaging technique is known as a promising non-destructive way for detecting plants diseases and pests. In most previous studies, the utilization of the whole spectrum or a large number of bands as well as the complexity of model structure severely hampers the application of the technique in practice. If a detection system can be established with a few bands and a relatively simple logic, it would be of great significance for application. This study established a method for identifying and discriminating three commonly occurring diseases and pests of wheat, i.e., powdery mildew, yellow rust and aphid with a few specific bands. Through a comprehensive spectral analysis, only three bands at 570, 680 and 750 nm were selected. A novel vegetation index namely Ratio Triangular Vegetation Index (RTVI) was developed for detecting anomalous areas on leaves. Then, the Support Vector Machine (SVM) method was applied to construct the discrimination model based on the spectral ratio analysis. The validating results suggested that the proposed method with only three spectral bands achieved a promising accuracy with the Overall Accuracy (OA) of 83%. With three bands from the hyperspectral imaging data, the three wheat diseases and pests were successfully detected and discriminated. A stepwise strategy including background removal, damage lesions recognition and stresses discrimination was proposed. The present work can provide a basis for the design of low cost and smart instruments for disease and pest detection.     

Plant Disease Severity Estimated Visually,by Digital Photography and Image Analysis,and by Hyperspectral Imaging

Reliable, precise and accurate estimates of disease severity are important for predicting yield loss, monitoring and forecasting epidemics, for assessing crop germplasm for disease resistance, and for understanding fundamental biological processes including co-evolution. Disease assessments that are inaccurate and/or imprecise might lead to faulty conclusions being drawn from the data, which in turn can lead to incorrect actions being taken in disease management decisions. Plant disease can be quantified in several different ways. This review considers plant disease severity assessment at the scale of individual plant parts or plants, and describes our current understanding of the sources and causes of assessment error, a better understanding of which is required before improvements can be targeted. The review also considers how these can be identified using various statistical tools. Indeed, great strides have been made in the last thirty years in identifying the sources of assessment error inherent to visual rating, and this review highlights ways that assessment errors can be reduced—particularly by training raters or using assessment aids. Lesion number in relation to area infected is known to influence accuracy and precision of visual estimates—the greater the number of lesions for a given area infected results in more overestimation. Furthermore, there is a widespread tendency to overestimate disease severity at low severities (<10%). Both interrater and intrarater reliability can be variable, particularly if training or rating aids are not used. During the last eighty years acceptable accuracy and precision of visual disease assessments have often been achieved using disease scales, particularly because of the time they allegedly save, and the ease with which they can be learned, but recent work suggests there can be some disadvantages to their use. This review considers new technologies that offer opportunity to assess disease with greater objectivity (reliability, precision, and accuracy). One of these, visible light photography and digital image analysis has been increasingly used over the last thirty years, as software has become more sophisticated and user-friendly. Indeed, some studies have produced very accurate estimates of disease using image analysis. In contrast, hyperspectral imagery is relatively recent and has not been widely applied in plant pathology. Nonetheless, it offers interesting and potentially discerning opportunities to assess disease. As plant disease assessment becomes better understood, it is against the backdrop of concepts of reliability, precision and accuracy (and agreement) in plant pathology and measurement science. This review briefly describes these concepts in relation to plant disease assessment. Various advantages and disadvantages of the different approaches to disease assessment are described. For each assessment method some future research priorities are identified that would be of value in better understanding the theory of disease assessment, as it applies to improving and fully realizing the potential of image analysis and hyperspectral imagery.     

Identification of Metal Oxide Nanoparticles in Histological Samples by Enhanced Darkfield Microscopy and Hyperspectral Mapping

Nanomaterials are increasingly prevalent throughout industry, manufacturing, and biomedical research. The need for tools and techniques that aid in the identification, localization, and characterization of nanoscale materials in biological samples is on the rise. Currently available methods, such as electron microscopy, tend to be resource-intensive, making their use prohibitive for much of the research community. Enhanced darkfield microscopy complemented with a hyperspectral imaging system may provide a solution to this bottleneck by enabling rapid and less expensive characterization of nanoparticles in histological samples. This method allows for high-contrast nanoscale imaging as well as nanomaterial identification. For this technique, histological tissue samples are prepared as they would be for light-based microscopy. First, positive control samples are analyzed to generate the reference spectra that will enable the detection of a material of interest in the sample. Negative controls without the material of interest are also analyzed in order to improve specificity (reduce false positives). Samples can then be imaged and analyzed using methods and software for hyperspectral microscopy or matched against these reference spectra in order to provide maps of the location of materials of interest in a sample. The technique is particularly well-suited for materials with highly unique reflectance spectra, such as noble metals, but is also applicable to other materials, such as semi-metallic oxides. This technique provides information that is difficult to acquire from histological samples without the use of electron microscopy techniques, which may provide higher sensitivity and resolution, but are vastly more resource-intensive and time-consuming than light microscopy.     

Detection and Discrimination of Tea Plant Stresses Based on Hyperspectral Imaging Technique at a Canopy Level

Tea plant stresses threaten the quality of tea seriously. The technology corresponding to the fast detection and differentiation of stresses is of great significance for plant protection in tea plantation. In recent years, hyperspectral imaging technology has shown great potential in detecting and differentiating plant diseases, pests and some other stresses at the leaf level. However, the lack of studies at canopy level hampers the detection of tea plant stresses at a larger scale. In this study, based on the canopy-level hyperspectral imaging data, the methods for identifying and differentiating the three commonly occurred tea stresses (i.e., the tea leafhopper, anthrax and sun burn) were studied. To account for the complexity of the canopy scenario, a stepwise detecting strategy was proposed that includes the process of background removal, identification of damaged areas and discrimination of stresses. Firstly, combining the successive projection algorithm (SPA) spectral analysis and K-means cluster analysis, the background and overexposed non-plant regions were removed from the image. Then, a rigorous sensitivity analysis and optimization were performed on various forms of spectral features, which yielded optimal features for detecting damaged areas (i.e., YSV, Area, GI, CARI and NBNDVI) and optimal features for stresses discrimination (i.e., MCARI, CI, LCI, RARS, TCI and VOG). Based on this information, the models for identifying damaged areas and those models for discriminating different stresses were established using K-nearest neighbor (KNN), Random Forest (RF) and Fisher discriminant analysis. The identification model achieved an accuracy over 95%, and the discrimination model achieved an accuracy over 93% for all stresses. The results suggested the feasibility of stress detection and differentiation using canopy-level hyperspectral imaging techniques, and indicated the potential for its extension over large areas.     

Hyperspectral imaging as a diagnostic tool to differentiate between amalgam tattoos and other dark pigmented intraoral lesions

The goal of this project is to identify any in-depth benefits and drawbacks in the diagnosis of amalgam tattoos and other pigmented intraoral lesions using hyperspectral imagery collected from amalgam tattoos, benign, and malignant melanocytic neoplasms. Software solutions capable of classifying pigmented lesions of the skin already exist, but conventional red, green and blue images may be reaching an upper limit in their performance. Emerging technologies, such as hyperspectral imaging (HSI) utilize more than a hundred, continuous data channels, while also collecting data in the infrared. A total of 18 paraffin-embedded human tissue specimens of dark pigmented intraoral lesions (including the lip) were analyzed using visible and near-infrared (VIS–NIR) hyperspectral imagery obtained from HE-stained histopathological slides. Transmittance data were collected between 450 and 900 nm using a snapshot camera mounted to a microscope with a halogen light source. VIS–NIR spectra collected from different specimens, such as melanocytic cells and other tissues (eg, epithelium), produced distinct and diagnostic spectra that were used to identify these materials in several regions of interest, making it possible to distinguish between intraoral amalgam tattoos (intramucosal metallic foreign bodies) and melanocytic lesions of the intraoral mucosa and the lip (each with P < .01 using the independent t test). HSI is presented as a diagnostic tool for the rapidly growing field of digital pathology. In this preliminary study, amalgam tattoos were reliably differentiated from melanocytic lesions of the oral cavity and the lip.     

Hyperspectral imaging and characterization of allergic contact dermatitis in the short‐wave infrared

Short‐wave infrared hyperspectral imaging is applied to diagnose and monitor a case of allergic contact dermatitis (ACD) due to poison ivy exposure in one subject. This approach directly demonstrates increased tissue fluid content in ACD lesional skin with a spectral signature that matches the spectral signature of intradermally injected normal saline. The best contrast between the affected and unaffected skin is achieved through a selection of specific wavelengths at 1070, 1340 and 1605 nm and combining them in a pseudo‐red‐green‐blue color space. An image derived from these wavelengths normalized to unaffected skin defines a “tissue fluid index” that may aid in the quantitative diagnosis and monitoring of ACD. Further clinical testing of this promising approach towards disease detection and monitoring with tissue fluid content quantification is warranted.