Spectral imaging in preclinical research and clinical pathology.

Spectral imaging methods are attracting increased interest from researchers and practitioners in basic science, pre-clinical and clinical arenas. A combination of better labeling reagents and better optics creates opportunities to detect and measure multiple parameters at the molecular and cellular level. These tools can provide valuable insights into the basic mechanisms of life, and yield diagnostic and prognostic information for clinical applications. There are many multispectral technologies available, each with its own advantages and limitations. This chapter will present an overview of the rationale for spectral imaging, and discuss the hardware, software and sample labeling strategies that can optimize its usefulness in clinical settings

Mapping Prosopis glandulosa (mesquite) invasion in the arid environment of South African using remote sensing techniques

A dissertation submitted to the School of Geography, Archaeology and Environmental Studies, Faculty of Science, University of the Witwatersrand, Johannesburg, in fulfilment of requirements for the degree of Master of Science in Environmental Sciences. Johannesburg, March 2016.Mapping Prosopis glandulosa (mesquite) invasion in the arid environment of South Africa using remote sensing techniques Mureriwa, Nyasha Abstract Decades after the first introduction of the Prosopis spp. (mesquite) to South Africa in the late 1800s for its benefits, the invasive nature of the species became apparent as its spread in regions of South Africa resulting in devastating effects to biodiversity, ecosystems and the socio-economic wellbeing of affected regions. Various control and management practices that include biological, physical, chemical and integrated methods have been tested with minimal success as compared to the rapid spread of the species. From previous studies, it has been noted that one of the reasons for the low success rates in mesquite control and management is a lack of sufficient information on the species invasion dynamic in relation to its very similar co-existing species. In order to bridge this gap in knowledge, vegetation species mapping techniques that use remote sensing methods need to be tested for the monitoring, detection and mapping of the species spread. Unlike traditional field survey methods, remote sensing techniques are better at monitoring vegetation as they can cover very large areas and are time-effective and cost-effective. Thus, the aim of this research was to examine the possibility of mapping and spectrally discriminating Prosopis glandulosa from its native co-existing species in semi-arid parts of South Africa using remote sensing methods. The specific objectives of the study were to investigate the spectral separability between Prosopis glandulosa and its co-existing species using field spectral data as well as to upscale the results to different satellites resolutions. Two machine learning algorithms (Random Forest (RF) and Support Vector Machines (SVM)) were also tested in the mapping processes. The first chapter of the study evaluated the spectral discrimination of Prosopis glandulosa from three other species (Acacia karoo, Acacia mellifera and Ziziphus mucronata) in the study area using in-situ spectroscopy in conjunction with the newly developed guided regularized random forest (GRRF) algorithm in identifying key wavelengths for multiclass classification. The GRRF algorithm was used as a method of reducing the problem of high dimensionality associated with hyperspectral data. Results showed that there was an increase in the accuracy of discrimination between the four species when the full set of 1825 wavelengths was used in classification (79.19%) as compared to the classification used by the 11 key wavelengths identified by GRRF (88.59%). Results obtained from the second chapter showed that it is possible to spatially discriminate mesquite from its co-existing acacia species and other general land-cover types at a 2 m resolution with overall accuracies of 86.59% for RF classification and 85.98% for SVM classification. The last part of the study tested the use of the more cost effective SPOT-6 imagery and the RF and SVM algorithms in mapping Prosopis glandulosa invasion and its co-existing indigenous species. The 6 m resolution analysis obtained accuracies of 78.46% for RF and 77.62% for SVM. Overall it was concluded that spatial and spectral discrimination of Prosopis glandulosa from its native co-existing species in semi-arid South Africa was possible with high accuracies through the use of (i) two high resolution, new generation sensors namely, WorldView-2 and SPOT-6; (ii) two robust classification algorithms specifically, RF and SVM and (iii) the newly developed GRRF algorithm for variable selection and reducing the high dimensionality problem associated with hyperspectral data. Some recommendations for future studies include the replication of this study on a larger scale in different invaded areas across the country as well as testing the robustness of the RF and SVM classifiers by making use of other machine learning algorithms and classification methods in species discrimination. Keywords: Prosopis glandulosa, field spectroscopy, cost effectiveness, Guided Regularised Random Forest, Support Vector Machines, Worldview-2, Spot-

Highly Multiplexed Single Cell In Situ Transcriptomic Analysis

abstract: Spatial resolved detection and quantification of ribonucleic acid (RNA) molecules in single cell is crucial for the understanding of inherent biological issues, like mechanism of gene regulation or the development and maintenance of cell fate. Conventional methods for single cell RNA profiling, like single-cell RNA sequencing (scRNA-seq) or single-molecule fluorescent in situ hybridization (smFISH), suffer either from the loss of spatial information or the low detection throughput. In order to advance single-cell analysis, new approaches need to be developed with the ability to perform high-throughput detection while preserving spatial information of the subcellular location of target RNA molecules. Novel approaches for highly multiplexed single cell in situ transcriptomic analysis were developed by our group to enable single-cell comprehensive RNA profiling in their native spatial contexts. Reiterative FISH was demonstrated to be able to detect >100 RNA species in single cell in situ, while more sophisticated approaches, consecutive FISH (C-FISH) and switchable fluorescent oligonucleotide based FISH (SFO-FISH), have the potential for whole transcriptome profiling at the single molecule sensitivity. The introduction of a cleavable fluorescent tyramide even enables sensitive RNA profiling in intact tissues with high throughput. These approaches will have wide applications in studies of systems biology, molecular diagnosis and targeted therapies.Dissertation/ThesisDoctoral Dissertation Chemistry 201

Highly Multiplexed Single Cell in situ Protein Analysis with Cleavable Fluorescent Probes

abstract: Measurements of different molecular species from single cells have the potential to reveal cell-to-cell variations, which are precluded by population-based measurements. An increasing percentage of researches have been focused on proteins, for its central roles in biological processes. Immunofluorescence (IF) has been a well-established protein analysis platform. To gain comprehensive insights into cell biology and diagnostic pathology, a crucial direction would be to increase the multiplexity of current single cell protein analysis technologies. An azide-based chemical cleavable linker has been introduced to design and synthesis novel fluorescent probes. These probes allow cyclic immunofluorescence staining which leads to the feasibility of highly multiplexed single cell in situ protein profiling. These highly multiplexed imaging-based platforms have the potential to quantify more than 100 protein targets in cultured cells and more than 50 protein targets in single cells in tissues. This approach has been successfully applied in formalin-fixed paraffin-embedded (FFPE) brain tissues. Multiplexed protein expression level results reveal neuronal heterogeneity in the human hippocampus.Dissertation/ThesisDoctoral Dissertation Chemistry 201

Hyperspectral Imaging for Landmine Detection

This PhD thesis aims at investigating the possibility to detect landmines using hyperspectral imaging. Using this technology, we are able to acquire at each pixel of the image spectral data in hundreds of wavelengths. So, at each pixel we obtain a reflectance spectrum that is used as fingerprint to identify the materials in each pixel, and mainly in our project help us to detect the presence of landmines. The proposed process works as follows: a preconfigured drone (hexarotor or octorotor) will carry the hyperspectral camera. This programmed drone is responsible of flying over the contaminated area in order to take images from a safe distance. Various image processing techniques will be used to treat the image in order to isolate the landmine from the surrounding. Once the presence of a mine or explosives is suspected, an alarm signal is sent to the base station giving information about the type of the mine, its location and the clear path that could be taken by the mine removal team in order to disarm the mine. This technology has advantages over the actually used techniques: • It is safer because it limits the need of humans in the searching process and gives the opportunity to the demining team to detect the mines while they are in a safe region. • It is faster. A larger area could be cleared in a single day by comparison with demining techniques • This technique can be used to detect at the same time objects other than mines such oil or minerals. First, a presentation of the problem of landmines that is expanding worldwide referring to some statistics from the UN organizations is provided. In addition, a brief presentation of different types of landmines is shown. Unfortunately, new landmines are well camouflaged and are mainly made of plastic in order to make their detection using metal detectors harder. A summary of all landmine detection techniques is shown to give an idea about the advantages and disadvantages of each technique. In this work, we give an overview of different projects that worked on the detection of landmines using hyperspectral imaging. We will show the main results achieved in this field and future work to be done in order to make this technology effective. Moreover, we worked on different target detection algorithms in order to achieve high probability of detection with low false alarm rate. We tested different statistical and linear unmixing based methods. In addition, we introduced the use of radial basis function neural networks in order to detect landmines at subpixel level. A comparative study between different detection methods will be shown in the thesis. A study of the effect of dimensionality reduction using principal component analysis prior to classification is also provided. The study shows the dependency between the two steps (feature extraction and target detection). The selection of target detection algorithm will define if feature extraction in previous phase is necessary. A field experiment has been done in order to study how the spectral signature of landmine will change depending on the environment in which the mine is planted. For this, we acquired the spectral signature of 6 types of landmines in different conditions: in Lab where specific source of light is used; in field where mines are covered by grass; and when mines are buried in soil. The results of this experiment are very interesting. The signature of two types of landmines are used in the simulations. They are a database necessary for supervised detection of landmines. Also we extracted some spectral characteristics of landmines that would help us to distinguish mines from background

Integrating optical imaging techniques for a novel approach to evaluate Siberian wild rye seed maturity

Advances in optical imaging technology using rapid and non-destructive methods have led to improvements in the efficiency of seed quality detection. Accurately timing the harvest is crucial for maximizing the yield of higher-quality Siberian wild rye seeds by minimizing excessive shattering during harvesting. This research applied integrated optical imaging techniques and machine learning algorithms to develop different models for classifying Siberian wild rye seeds based on different maturity stages and grain positions. The multi-source fusion of morphological, multispectral, and autofluorescence data provided more comprehensive information but also increases the performance requirements of the equipment. Therefore, we employed three filtering algorithms, namely minimal joint mutual information maximization (JMIM), information gain, and Gini impurity, and set up two control methods (feature union and no-filtering) to assess the impact of retaining only 20% of the features on the model performance. Both JMIM and information gain revealed autofluorescence and morphological features (CIELab A, CIELab B, hue and saturation), with these two filtering algorithms showing shorter run times. Furthermore, a strong correlation was observed between shoot length and morphological and autofluorescence spectral features. Machine learning models based on linear discriminant analysis (LDA), random forests (RF) and support vector machines (SVM) showed high performance (>0.78 accuracies) in classifying seeds at different maturity stages. Furthermore, it was found that there was considerable variation in the different grain positions at the maturity stage, and the K-means approach was used to improve the model performance by 5.8%-9.24%. In conclusion, our study demonstrated that feature filtering algorithms combined with machine learning algorithms offer high performance and low cost in identifying seed maturity stages and that the application of k-means techniques for inconsistent maturity improves classification accuracy. Therefore, this technique could be employed classification of seed maturity and superior physiological quality for Siberian wild rye seeds

Neuro-Fuzzy Based Intelligent Approaches to Nonlinear System Identification and Forecasting

Nearly three decades back nonlinear system identification consisted of several ad-hoc approaches, which were restricted to a very limited class of systems. However, with the advent of the various soft computing methodologies like neural networks and the fuzzy logic combined with optimization techniques, a wider class of systems can be handled at present. Complex systems may be of diverse characteristics and nature. These systems may be linear or nonlinear, continuous or discrete, time varying or time invariant, static or dynamic, short term or long term, central or distributed, predictable or unpredictable, ill or well defined. Neurofuzzy hybrid modelling approaches have been developed as an ideal technique for utilising linguistic values and numerical data. This Thesis is focused on the development of advanced neurofuzzy modelling architectures and their application to real case studies. Three potential requirements have been identified as desirable characteristics for such design: A model needs to have minimum number of rules; a model needs to be generic acting either as Multi-Input-Single-Output (MISO) or Multi-Input-Multi-Output (MIMO) identification model; a model needs to have a versatile nonlinear membership function. Initially, a MIMO Adaptive Fuzzy Logic System (AFLS) model which incorporates a prototype defuzzification scheme, while utilising an efficient, compared to the Takagi–Sugeno–Kang (TSK) based systems, fuzzification layer has been developed for the detection of meat spoilage using Fourier transform infrared (FTIR) spectroscopy. The identification strategy involved not only the classification of beef fillet samples in their respective quality class (i.e. fresh, semi-fresh and spoiled), but also the simultaneous prediction of their associated microbiological population directly from FTIR spectra. In the case of AFLS, the number of memberships for each input variable was directly associated to the number of rules, hence, the “curse of dimensionality” problem was significantly reduced. Results confirmed the advantage of the proposed scheme against Adaptive Neurofuzzy Inference System (ANFIS), Multilayer Perceptron (MLP) and Partial Least Squares (PLS) techniques used in the same case study. In the case of MISO systems, the TSK based structure, has been utilized in many neurofuzzy systems, like ANFIS. At the next stage of research, an Adaptive Fuzzy Inference Neural Network (AFINN) has been developed for the monitoring the spoilage of minced beef utilising multispectral imaging information. This model, which follows the TSK structure, incorporates a clustering pre-processing stage for the definition of fuzzy rules, while its final fuzzy rule base is determined by competitive learning. In this specific case study, AFINN model was also able to predict for the first time in the literature, the beef’s temperature directly from imaging information. Results again proved the superiority of the adopted model. By extending the line of research and adopting specific design concepts from the previous case studies, the Asymmetric Gaussian Fuzzy Inference Neural Network (AGFINN) architecture has been developed. This architecture has been designed based on the above design principles. A clustering preprocessing scheme has been applied to minimise the number of fuzzy rules. AGFINN incorporates features from the AFLS concept, by having the same number of rules as well as fuzzy memberships. In spite of the extensive use of the standard symmetric Gaussian membership functions, AGFINN utilizes an asymmetric function acting as input linguistic node. Since the asymmetric Gaussian membership function’s variability and flexibility are higher than the traditional one, it can partition the input space more effectively. AGFINN can be built either as an MISO or as an MIMO system. In the MISO case, a TSK defuzzification scheme has been implemented, while two different learning algorithms have been implemented. AGFINN has been tested on real datasets related to electricity price forecasting for the ISO New England Power Distribution System. Its performance was compared against a number of alternative models, including ANFIS, AFLS, MLP and Wavelet Neural Network (WNN), and proved to be superior. The concept of asymmetric functions proved to be a valid hypothesis and certainly it can find application to other architectures, such as in Fuzzy Wavelet Neural Network models, by designing a suitable flexible wavelet membership function. AGFINN’s MIMO characteristics also make the proposed architecture suitable for a larger range of applications/problems

A review of neural networks in plant disease detection using hyperspectral data

© 2018 China Agricultural University This paper reviews advanced Neural Network (NN) techniques available to process hyperspectral data, with a special emphasis on plant disease detection. Firstly, we provide a review on NN mechanism, types, models, and classifiers that use different algorithms to process hyperspectral data. Then we highlight the current state of imaging and non-imaging hyperspectral data for early disease detection. The hybridization of NN-hyperspectral approach has emerged as a powerful tool for disease detection and diagnosis. Spectral Disease Index (SDI) is the ratio of different spectral bands of pure disease spectra. Subsequently, we introduce NN techniques for rapid development of SDI. We also highlight current challenges and future trends of hyperspectral data

Ash Tree Identification Based on the Integration of Hyperspectral Imagery and High-density Lidar Data

Monitoring and management of ash trees has become particularly important in recent years due to the heightened risk of attack from the invasive pest, the emerald ash borer (EAB). However, distinguishing ash from other deciduous trees can be challenging. Both hyperspectral imagery and Light detection and ranging (LiDAR) data are two valuable data sources that are often used for tree species classification. Hyperspectral imagery measures detailed spectral reflectance related to the biochemical properties of vegetation, while LiDAR data measures the three-dimensional structure of tree crowns related to morphological characteristics. Thus, the accuracy of vegetation classification may be improved by combining both techniques. Therefore, the objective of this research is to integrate hyperspectral imagery and LiDAR data for improving ash tree identification. Specifically, the research aims include: 1) using LiDAR data for individual tree crowns segmentation; 2) using hyperspectral imagery for extraction of relative pure crown spectra; 3) fusing hyperspectral and LiDAR data for ash tree identification. It is expected that the classification accuracy of ash trees will be significantly improved with the integration of hyperspectral and LiDAR techniques. Analysis results suggest that, first, 3D crown structures of individual trees can be reconstructed using a set of generalized geometric models which optimally matched LiDAR-derived raster image, and crown widths can be further estimated using tree height and shape-related parameters as independent variables and ground measurement of crown widths as dependent variables. Second, with constrained linear spectral mixture analysis method, the fractions of all materials within a pixel can be extracted, and relative pure crown-scale spectra can be further calculated using illuminated-leaf fraction as weighting factors for tree species classification. Third, both crown shape index (SI) and coefficient of variation (CV) can be extracted from LiDAR data as invariant variables in tree’s life cycle, and improve ash tree identification by integrating with pixel-weighted crown spectra. Therefore, three major contributions of this research have been made in the field of tree species classification:1) the automatic estimation of individual tree crown width from LiDAR data by combining a generalized geometric model and a regression model, 2) the computation of relative pure crown-scale spectral reflectance using a pixel-weighting algorithm for tree species classification, 3) the fusion of shape-related structural features and pixel-weighted crown-scale spectral features for improving of ash tree identification