Multiplexed single-cell in situ RNA analysis by reiterative hybridization

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Description
Currently, quantification of single cell RNA species in their natural contexts is restricted due to the little number of parallel analysis. Through this, we identify a method to increase the multiplexing capacity of RNA analysis for single cells in situ.…

Currently, quantification of single cell RNA species in their natural contexts is restricted due to the little number of parallel analysis. Through this, we identify a method to increase the multiplexing capacity of RNA analysis for single cells in situ. Initially, RNA transcripts are found by using fluorescence in situ hybridization (FISH). Once imaging and data storage is completed, the fluorescence signal is detached through photobleaching. By doing so, the FISH is reinitiated to detect other RNA species residing in the same cell. After reiterative cycles of hybridization, imaging and photobleaching, the identities, positions and copy numbers of a huge amount of varied RNA species can be computed in individual cells in situ. Through this approach, we have evaluated seven different transcripts in single HeLa cells with five reiterative RNA FISH cycles. This method has the ability to detect over 100 varied RNA species in single cells in situ, which can be further applied in studies of systems biology, molecular diagnosis and targeted therapies.
Date Created
2016-12
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Use of cleavable fluorescent antibodies for highly multiplexed single cell in situ protein analysis

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Description
The ability to profile proteins allows us to gain a deeper understanding of organization, regulation, and function of different biological systems. Many technologies are currently being used in order to accurately perform the protein profiling. Some of these technologies include…

The ability to profile proteins allows us to gain a deeper understanding of organization, regulation, and function of different biological systems. Many technologies are currently being used in order to accurately perform the protein profiling. Some of these technologies include mass spectrometry, microarray based analysis, and fluorescence microscopy. Deeper analysis of these technologies have demonstrated limitations which have taken away from either the efficiency or the accuracy of the results. The objective of this project was to develop a technology in which highly multiplexed single cell in situ protein analysis can be completed in a comprehensive manner without the loss of the protein targets. This was accomplished in the span of 3 steps which is referred to as the immunofluorescence cycle. Antibodies with attached fluorophores with the help of novel azide-based cleavable linker are used to detect protein targets. Fluorescence imaging and data storage procedures are done on the targets and then the fluorophores are cleaved from the antibodies without the loss of the protein targets. Continuous cycles of the immunofluorescence procedure can help create a comprehensive and quantitative profile of the protein. The development of such a technique will not only help us understand biological systems such as solid tumor, brain tissues, and developing embryos. But it will also play a role in real-world applications such as signaling network analysis, molecular diagnosis and cellular targeted therapies.
Date Created
2016-12
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Reconciling the Differences Between a Bottom-Up and Inverse-Estimated FFCO2 Emissions Estimate in a Large U.S. Urban Area

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Description

The INFLUX experiment has taken multiple approaches to estimate the carbon dioxide (CO2) flux in a domain centered on the city of Indianapolis, Indiana. One approach, Hestia, uses a bottom-up technique relying on a mixture of activity data, fuel statistics,…

The INFLUX experiment has taken multiple approaches to estimate the carbon dioxide (CO2) flux in a domain centered on the city of Indianapolis, Indiana. One approach, Hestia, uses a bottom-up technique relying on a mixture of activity data, fuel statistics, direct flux measurement and modeling algorithms. A second uses a Bayesian atmospheric inverse approach constrained by atmospheric CO2 measurements and the Hestia emissions estimate as a prior CO2 flux. The difference in the central estimate of the two approaches comes to 0.94 MtC (an 18.7% difference) over the eight-month period between September 1, 2012 and April 30, 2013, a statistically significant difference at the 2-sigma level. Here we explore possible explanations for this apparent discrepancy in an attempt to reconcile the flux estimates. We focus on two broad categories: 1) biases in the largest of bottom-up flux contributions and 2) missing CO2 sources. Though there is some evidence for small biases in the Hestia fossil fuel carbon dioxide (FFCO2) flux estimate as an explanation for the calculated difference, we find more support for missing CO2 fluxes, with biological respiration the largest of these. Incorporation of these differences bring the Hestia bottom-up and the INFLUX inversion flux estimates into statistical agreement and are additionally consistent with wintertime measurements of atmospheric 14CO2. We conclude that comparison of bottom-up and top-down approaches must consider all flux contributions and highlight the important contribution to urban carbon budgets of animal and biotic respiration. Incorporation of missing CO2 fluxes reconciles the bottom-up and inverse-based approach in the INFLUX domain.

Date Created
2017-08-03
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Optimizing the Spatial Resolution for Urban CO2 Flux Studies Using the Shannon Entropy

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Description

The ‘Hestia Project’ uses a bottom-up approach to quantify fossil fuel CO2(FFCO2) emissions spatially at the building/street level and temporally at the hourly level. Hestia FFCO2 emissions are provided in the form of a group of sector-specific vector layers with…

The ‘Hestia Project’ uses a bottom-up approach to quantify fossil fuel CO2(FFCO2) emissions spatially at the building/street level and temporally at the hourly level. Hestia FFCO2 emissions are provided in the form of a group of sector-specific vector layers with point, line, and polygon sources to support carbon cycle science and climate policy. Application to carbon cycle science, in particular, requires regular gridded data in order to link surface carbon fluxes to atmospheric transport models. However, the heterogeneity and complexity of FFCO2 sources within regular grids is sensitive to spatial resolution. From the perspective of a data provider, we need to find a balance between resolution and data volume so that the gridded data product retains the maximum amount of information content while maintaining an efficient data volume.

The Shannon entropy determines the minimum bits that are needed to encode an information source and can serve as a metric for the effective information content. In this paper, we present an analysis of the Shannon entropy of gridded FFCO2 emissions with varying resolutions in four Hestia study areas, and find: (1) the Shannon entropy increases with smaller grid resolution until it reaches a maximum value (the max-entropy resolution); (2) total emissions (the sum of several sector-specific emission fields) show a finer max-entropy resolution than each of the sector-specific fields; (3) the residential emissions show a finer max-entropy resolution than the commercial emissions; (4) the max-entropy resolution of the onroad emissions grid is closely correlated to the density of the road network. These findings suggest that the Shannon entropy can detect the information effectiveness of the spatial resolution of gridded FFCO2 emissions. Hence, the resolution-entropy relationship can be used to assist in determining an appropriate spatial resolution for urban CO2 flux studies. We conclude that the optimal spatial resolution for providing Hestia total FFCO2 emissions products is centered around 100 m, at which the FFCO2 emissions data can not only fully meet the requirement of urban flux integration, but also be effectively used in understanding the relationships between FFCO2 emissions and various social-economic variables at the U.S. census block group level.

Date Created
2017-05-19
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3D - Patch Based Machine Learning Systems for Alzheimer’s Disease classification via 18F-FDG PET Analysis

Description
Alzheimer’s disease (AD), is a chronic neurodegenerative disease that usually starts slowly and gets worse over time. It is the cause of 60% to 70% of cases of dementia. There is growing interest in identifying brain image biomarkers that hel…

Alzheimer’s disease (AD), is a chronic neurodegenerative disease that usually starts slowly and gets worse over time. It is the cause of 60% to 70% of cases of dementia. There is growing interest in identifying brain image biomarkers that help evaluate AD risk pre-symptomatically. High-dimensional non-linear pattern classification methods have been applied to structural magnetic resonance images (MRI’s) and used to discriminate between clinical groups in Alzheimers progression. Using Fluorodeoxyglucose (FDG) positron emission tomography (PET) as the pre- ferred imaging modality, this thesis develops two independent machine learning based patch analysis methods and uses them to perform six binary classification experiments across different (AD) diagnostic categories. Specifically, features were extracted and learned using dimensionality reduction and dictionary learning & sparse coding by taking overlapping patches in and around the cerebral cortex and using them as fea- tures. Using AdaBoost as the preferred choice of classifier both methods try to utilize 18F-FDG PET as a biological marker in the early diagnosis of Alzheimer’s . Addi- tional we investigate the involvement of rich demographic features (ApoeE3, ApoeE4 and Functional Activities Questionnaires (FAQ)) in classification. The experimental results on Alzheimer’s Disease Neuroimaging initiative (ADNI) dataset demonstrate the effectiveness of both the proposed systems. The use of 18F-FDG PET may offer a new sensitive biomarker and enrich the brain imaging analysis toolset for studying the diagnosis and prognosis of AD.
Date Created
2017
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Deep Learning based Classification of FDG-PET Data for Alzheimer's Disease

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Description
Alzheimer’s Disease (AD), a neurodegenerative disease is a progressive disease that affects the brain gradually with time and worsens. Reliable and early diagnosis of AD and its prodromal stages (i.e. Mild Cognitive Impairment(MCI)) is essential. Fluorodeoxyglucose (FDG) positron emission tomography…

Alzheimer’s Disease (AD), a neurodegenerative disease is a progressive disease that affects the brain gradually with time and worsens. Reliable and early diagnosis of AD and its prodromal stages (i.e. Mild Cognitive Impairment(MCI)) is essential. Fluorodeoxyglucose (FDG) positron emission tomography (PET) measures the decline in the regional cerebral metabolic rate for glucose, offering a reliable metabolic biomarker even on presymptomatic AD patients. PET scans provide functional information that is unique and unavailable using other types of imaging. The computational efficacy of FDG-PET data alone, for the classification of various Alzheimer’s Diagnostic categories (AD, MCI (LMCI, EMCI), Control) has not been studied. This serves as motivation to correctly classify the various diagnostic categories using FDG-PET data. Deep learning has recently been applied to the analysis of structural and functional brain imaging data. This thesis is an introduction to a deep learning based classification technique using neural networks with dimensionality reduction techniques to classify the different stages of AD based on FDG-PET image analysis.

This thesis develops a classification method to investigate the performance of FDG-PET as an effective biomarker for Alzheimer's clinical group classification. This involves dimensionality reduction using Probabilistic Principal Component Analysis on max-pooled data and mean-pooled data, followed by a Multilayer Feed Forward Neural Network which performs binary classification. Max pooled features result into better classification performance compared to results on mean pooled features. Additionally, experiments are done to investigate if the addition of important demographic features such as Functional Activities Questionnaire(FAQ), gene information helps improve performance. Classification results indicate that our designed classifiers achieve competitive results, and better with the additional of demographic features.
Date Created
2017
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A Sparse Voxel Octree-Based Framework for Computing Solar Radiation Using 3D City Models

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Description

An effective three-dimensional (3D) data representation is required to assess the spatial distribution of the photovoltaic potential over urban building roofs and facades using 3D city models. Voxels have long been used as a spatial data representation, but practical applications…

An effective three-dimensional (3D) data representation is required to assess the spatial distribution of the photovoltaic potential over urban building roofs and facades using 3D city models. Voxels have long been used as a spatial data representation, but practical applications of the voxel representation have been limited compared with rasters in traditional two-dimensional (2D) geographic information systems (GIS). We propose to use sparse voxel octree (SVO) as a data representation to extend the GRASS GIS r.sun solar radiation model from 2D to 3D. The GRASS GIS r.sun model is nested in an SVO-based computing framework. The presented 3D solar radiation computing framework was applied to 3D building groups of different geometric complexities to demonstrate its efficiency and scalability. We presented a method to explicitly compute diffuse shading losses in r.sun, and found that diffuse shading losses can reduce up to 10% of the annual global radiation under clear sky conditions. Hence, diffuse shading losses are of significant importance especially in complex urban environments.

Date Created
2017-03-31
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Video2Vec: learning semantic spatio-temporal embedding for video representations

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Description
High-level inference tasks in video applications such as recognition, video retrieval, and zero-shot classification have become an active research area in recent years. One fundamental requirement for such applications is to extract high-quality features that maintain high-level information in the…

High-level inference tasks in video applications such as recognition, video retrieval, and zero-shot classification have become an active research area in recent years. One fundamental requirement for such applications is to extract high-quality features that maintain high-level information in the videos.

Many video feature extraction algorithms have been purposed, such as STIP, HOG3D, and Dense Trajectories. These algorithms are often referred to as “handcrafted” features as they were deliberately designed based on some reasonable considerations. However, these algorithms may fail when dealing with high-level tasks or complex scene videos. Due to the success of using deep convolution neural networks (CNNs) to extract global representations for static images, researchers have been using similar techniques to tackle video contents. Typical techniques first extract spatial features by processing raw images using deep convolution architectures designed for static image classifications. Then simple average, concatenation or classifier-based fusion/pooling methods are applied to the extracted features. I argue that features extracted in such ways do not acquire enough representative information since videos, unlike images, should be characterized as a temporal sequence of semantically coherent visual contents and thus need to be represented in a manner considering both semantic and spatio-temporal information.

In this thesis, I propose a novel architecture to learn semantic spatio-temporal embedding for videos to support high-level video analysis. The proposed method encodes video spatial and temporal information separately by employing a deep architecture consisting of two channels of convolutional neural networks (capturing appearance and local motion) followed by their corresponding Fully Connected Gated Recurrent Unit (FC-GRU) encoders for capturing longer-term temporal structure of the CNN features. The resultant spatio-temporal representation (a vector) is used to learn a mapping via a Fully Connected Multilayer Perceptron (FC-MLP) to the word2vec semantic embedding space, leading to a semantic interpretation of the video vector that supports high-level analysis. I evaluate the usefulness and effectiveness of this new video representation by conducting experiments on action recognition, zero-shot video classification, and semantic video retrieval (word-to-video) retrieval, using the UCF101 action recognition dataset.
Date Created
2016
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A computational approach to relative image aesthetics

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Description
Computational visual aesthetics has recently become an active research area. Existing state-of-art methods formulate this as a binary classification task where a given image is predicted to be beautiful or not. In many applications such as image retrieval and enhancement,…

Computational visual aesthetics has recently become an active research area. Existing state-of-art methods formulate this as a binary classification task where a given image is predicted to be beautiful or not. In many applications such as image retrieval and enhancement, it is more important to rank images based on their aesthetic quality instead of binary-categorizing them. Furthermore, in such applications, it may be possible that all images belong to the same category. Hence determining the aesthetic ranking of the images is more appropriate. To this end, a novel problem of ranking images with respect to their aesthetic quality is formulated in this work. A new data-set of image pairs with relative labels is constructed by carefully selecting images from the popular AVA data-set. Unlike in aesthetics classification, there is no single threshold which would determine the ranking order of the images across the entire data-set.

This problem is attempted using a deep neural network based approach that is trained on image pairs by incorporating principles from relative learning. Results show that such relative training procedure allows the network to rank the images with a higher accuracy than a state-of-art network trained on the same set of images using binary labels. Further analyzing the results show that training a model using the image pairs learnt better aesthetic features than training on same number of individual binary labelled images.

Additionally, an attempt is made at enhancing the performance of the system by incorporating saliency related information. Given an image, humans might fixate their vision on particular parts of the image, which they might be subconsciously intrigued to. I therefore tried to utilize the saliency information both stand-alone as well as in combination with the global and local aesthetic features by performing two separate sets of experiments. In both the cases, a standard saliency model is chosen and the generated saliency maps are convoluted with the images prior to passing them to the network, thus giving higher importance to the salient regions as compared to the remaining. Thus generated saliency-images are either used independently or along with the global and the local features to train the network. Empirical results show that the saliency related aesthetic features might already be learnt by the network as a sub-set of the global features from automatic feature extraction, thus proving the redundancy of the additional saliency module.
Date Created
2016
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A unified framework based on convolutional neural networks for interpreting carotid intima-media thickness videos

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Description
Cardiovascular disease (CVD) is the leading cause of mortality yet largely preventable, but the key to prevention is to identify at-risk individuals before adverse events. For predicting individual CVD risk, carotid intima-media thickness (CIMT), a noninvasive ultrasound method, has proven…

Cardiovascular disease (CVD) is the leading cause of mortality yet largely preventable, but the key to prevention is to identify at-risk individuals before adverse events. For predicting individual CVD risk, carotid intima-media thickness (CIMT), a noninvasive ultrasound method, has proven to be valuable, offering several advantages over CT coronary artery calcium score. However, each CIMT examination includes several ultrasound videos, and interpreting each of these CIMT videos involves three operations: (1) select three enddiastolic ultrasound frames (EUF) in the video, (2) localize a region of interest (ROI) in each selected frame, and (3) trace the lumen-intima interface and the media-adventitia interface in each ROI to measure CIMT. These operations are tedious, laborious, and time consuming, a serious limitation that hinders the widespread utilization of CIMT in clinical practice. To overcome this limitation, this paper presents a new system to automate CIMT video interpretation. Our extensive experiments demonstrate that the suggested system significantly outperforms the state-of-the-art methods. The superior performance is attributable to our unified framework based on convolutional neural networks (CNNs) coupled with our informative image representation and effective post-processing of the CNN outputs, which are uniquely designed for each of the above three operations.
Date Created
2016
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