| 1. |
- Matuszewski, Damian J., et al.
(författare)
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Image-Based Detection of Patient-Specific Drug-Induced Cell-Cycle Effects in Glioblastoma
- 2018
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Ingår i: SLAS Discovery. - : Elsevier BV. - 2472-5560 .- 2472-5552. ; 23:10, s. 1030-1039
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Tidskriftsartikel (refereegranskat)abstract
- Image-based analysis is an increasingly important tool to characterize the effect of drugs in large-scale chemical screens. Herein, we present image and data analysis methods to investigate population cell-cycle dynamics in patient-derived brain tumor cells. Images of glioblastoma cells grown in multiwell plates were used to extract per-cell descriptors, including nuclear DNA content. We reduced the DNA content data from per-cell descriptors to per-well frequency distributions, which were used to identify compounds affecting cell-cycle phase distribution. We analyzed cells from 15 patient cases representing multiple subtypes of glioblastoma and searched for clusters of cell-cycle phase distributions characterizing similarities in response to 249 compounds at 11 doses. We show that this approach applied in a blind analysis with unlabeled substances identified drugs that are commonly used for treating solid tumors as well as other compounds that are well known for inducing cell-cycle arrest. Redistribution of nuclear DNA content signals is thus a robust metric of cell-cycle arrest in patient-derived glioblastoma cells.
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| 2. |
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| 3. |
- Blamey, Ben, et al.
(författare)
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Rapid development of cloud-native intelligent data pipelines for scientific data streams using the HASTE Toolkit
- 2021
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Ingår i: GigaScience. - : Oxford University Press. - 2047-217X. ; 10:3, s. 1-14
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Large streamed datasets, characteristic of life science applications, are often resource-intensive to process, transport and store. We propose a pipeline model, a design pattern for scientific pipelines, where an incoming stream of scientific data is organized into a tiered or ordered "data hierarchy". We introduce the HASTE Toolkit, a proof-of-concept cloud-native software toolkit based on this pipeline model, to partition and prioritize data streams to optimize use of limited computing resources.FINDINGS: In our pipeline model, an "interestingness function" assigns an interestingness score to data objects in the stream, inducing a data hierarchy. From this score, a "policy" guides decisions on how to prioritize computational resource use for a given object. The HASTE Toolkit is a collection of tools to adopt this approach. We evaluate with 2 microscopy imaging case studies. The first is a high content screening experiment, where images are analyzed in an on-premise container cloud to prioritize storage and subsequent computation. The second considers edge processing of images for upload into the public cloud for real-time control of a transmission electron microscope.CONCLUSIONS: Through our evaluation, we created smart data pipelines capable of effective use of storage, compute, and network resources, enabling more efficient data-intensive experiments. We note a beneficial separation between scientific concerns of data priority, and the implementation of this behaviour for different resources in different deployment contexts. The toolkit allows intelligent prioritization to be `bolted on' to new and existing systems - and is intended for use with a range of technologies in different deployment scenarios.
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| 4. |
- Colomb-Delsuc, Mathieu, et al.
(författare)
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Assessment of the percentage of full recombinant adeno-associated virus particles in a gene therapy drug using CryoTEM
- 2022
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Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 17:6
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Tidskriftsartikel (refereegranskat)abstract
- In spite of continuous development of gene therapy vectors with thousands of drug candidates in clinical drug trials there are only a small number approved on the market today stressing the need to have characterization methods to assist in the validation of the drug development process. The level of packaging of the vector capsids appears to play a critical role in immunogenicity, hence an objective quantitative method assessing the content of particles containing a genome is an essential quality measurement. As transmission electron microscopy (TEM) allows direct visualization of the particles present in a specimen, it naturally seems as the most intuitive method of choice for characterizing recombinant adeno-associated virus (rAAV) particle packaging. Negative stain TEM (nsTEM) is an established characterization method for analysing the packaging of viral vectors. It has however shown limitations in terms of reliability. To overcome this drawback, we propose an analytical method based on CryoTEM that unambiguously and robustly determines the percentage of filled particles in an rAAV sample. In addition, we show that at a fixed number of vector particles the portion of filled particles correlates well with the potency of the drug. The method has been validated according to the ICH Q2 (R1) guidelines and the components investigated during the validation are presented in this study. The reliability of nsTEM as a method for the assessment of filled particles is also investigated along with a discussion about the origin of the observed variability of this method.
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| 5. |
- Gupta, Ankit
(författare)
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Adapting Deep Learning for Microscopy: Interaction, Application, and Validation
- 2023
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Microscopy is an integral technique in biology to study the fundamental components of life visually. Digital microscopy and automation have enabled biologists to conduct faster and larger-scale experiments with a sharp increase in the data generated. Microscopy images contain rich but sparse information, as typically, only small regions in the images are relevant for further study. Image analysis is a crucial tool for biologists in the objective interpretation and extraction of quantitative measurements from microscopy data. Recently, deep learning techniques have shown superior performance in various image analysis tasks. The models learn feature representations from the data by optimizing for a task. However, the techniques require a significant amount of annotated data to perform well. Domain experts are required to annotate microscopy data, making it expensive and time-consuming. The models offer no insight into their prediction, and the learned features are not directly interpretable. This poses challenges to the reliable utilization of the technique in high-trust applications such as drug discovery or disease detection. High data variability in microscopy and poor generalization performance of deep learning models further increase the difficulty in general usage of the technique. The work in this thesis presents frameworks and methods to solve the practical challenges of applying deep learning in microscopy. The application-specific evaluation approaches were presented to validate the approaches, aiming to increase trust in the system. The major contributions of this work are as follows. Papers I and III present human-in-the-loop frameworks for quick adaption of deep learning to new data and for improving models' performance based on human input in visual explanations provided by the model, respectively. Paper II proposes a template-matching approach to improve user interactions in the framework proposed in Paper I. Papers III and IV present architectural modifications in the deep learning models proposed for better visual explanation and image-to-image translation, respectively. Papers IV and V present biologically relevant evaluations of approaches, i.e., analysis of the deep learning models in relation to the biological task.This thesis is aimed towards better utilization and adaptation of the DL methods and techniques to the microscopy data. We show that the annotation burden for the user can be significantly reduced by intuitive annotation frameworks and using contemporary deep-learning paradigms. We further propose architectural modifications in the models to adapt to the requirements and demonstrate the utility of application-specific analysis in microscopy.
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| 6. |
- Gupta, Anindya, et al.
(författare)
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Detection of pulmonary micronodules in computed tomography images and false positive reduction using 3D convolutional neural networks
- 2020
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Ingår i: International journal of imaging systems and technology (Print). - : Wiley. - 0899-9457 .- 1098-1098. ; 30:2, s. 327-339
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Tidskriftsartikel (refereegranskat)abstract
- Manual detection of small uncalcified pulmonary nodules (diameter <4 mm) in thoracic computed tomography (CT) scans is a tedious and error‐prone task. Automatic detection of disperse micronodules is, thus, highly desirable for improved characterization of the fatal and incurable occupational pulmonary diseases. Here, we present a novel computer‐assisted detection (CAD) scheme specifically dedicated to detect micronodules. The proposed scheme consists of a candidate‐screening module and a false positive (FP) reduction module. The candidate‐screening module is initiated by a lung segmentation algorithm and is followed by a combination of 2D/3D features‐based thresholding parameters to identify plausible micronodules. The FP reduction module employs a 3D convolutional neural network (CNN) to classify each identified candidate. It automatically encodes the discriminative representations by exploiting the volumetric information of each candidate. A set of 872 micro‐nodules in 598 CT scans marked by at least two radiologists are extracted from the Lung Image Database Consortium and Image Database Resource Initiative to test our CAD scheme. The CAD scheme achieves a detection sensitivity of 86.7% (756/872) with only 8 FPs/scan and an AUC of 0.98. Our proposed CAD scheme efficiently identifies micronodules in thoracic scans with only a small number of FPs. Our experimental results provide evidence that the automatically generated features by the 3D CNN are highly discriminant, thus making it a well‐suited FP reduction module of a CAD scheme.
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| 7. |
- Gupta, Ankit, et al.
(författare)
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Efficient High-Resolution Template Matching with Vector Quantized Nearest Neighbour Fields
- 2024
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Ingår i: Pattern Recognition. - : Elsevier. - 0031-3203 .- 1873-5142. ; 151
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Tidskriftsartikel (refereegranskat)abstract
- Template matching is a fundamental problem in computer vision with applications in fields including object detection, image registration, and object tracking. Current methods rely on nearest-neighbour (NN) matching, where the query feature space is converted to NN space by representing each query pixel with its NN in the template. NN-based methods have been shown to perform better in occlusions, appearance changes, and non-rigid transformations; however, they scale poorly with high-resolution data and high feature dimensions. We present an NN-based method that efficiently reduces the NN computations and introduces filtering in the NN fields (NNFs). A vector quantization step is introduced before the NN calculation to represent the template with k features, and the filter response over the NNFs is used to compare the template and query distributions over the features. We show that state-of-the-art performance is achieved in low-resolution data, and our method outperforms previous methods at higher resolution.
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| 8. |
- Gupta, Ankit, et al.
(författare)
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Is brightfield all you need for MoA prediction?
- 2022
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Konferensbidrag (refereegranskat)abstract
- Fluorescence staining techniques, such as Cell Painting, together with fluorescence microscopy have proven invaluable for visualizing and quantifying the effects that drugs and other perturbations have on cultured cells. However, fluorescence microscopy is expensive, time-consuming, and labor-intensive, and the stains applied can be cytotoxic, interfering with the activity under study. The simplest form of microscopy, brightfield microscopy, lacks these downsides, but the images produced have low contrast and the cellular compartments are difficult to discern. Nevertheless, by harnessing deep learning, these brightfield images may still be sufficient for various predictive purposes. In this study, we compared the predictive performance of models trained on fluorescence images to those trained on brightfield images for predicting the mechanism of action (MoA) of different drugs. We also extracted CellProfiler features from the fluorescence images and used them to benchmark the performance. Overall, we found comparable and correlated predictive performance for the two imaging modalities. This is promising for future studies of MoAs in time-lapse experiments.
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| 9. |
- Gupta, Ankit, et al.
(författare)
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SimSearch : A Human-in-The-Loop Learning Framework for Fast Detection of Regions of Interest in Microscopy Images
- 2022
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Ingår i: IEEE journal of biomedical and health informatics. - : Institute of Electrical and Electronics Engineers (IEEE). - 2168-2194 .- 2168-2208. ; 26:8, s. 4079-4089
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Tidskriftsartikel (refereegranskat)abstract
- Objective: Large-scale microscopy-based experiments often result in images with rich but sparse information content. An experienced microscopist can visually identify regions of interest (ROIs), but this becomes a cumbersome task with large datasets. Here we present SimSearch, a framework for quick and easy user-guided training of a deep neural model aimed at fast detection of ROIs in large-scale microscopy experiments. Methods: The user manually selects a small number of patches representing different classes of ROIs. This is followed by feature extraction using a pre-trained deep-learning model, and interactive patch selection pruning, resulting in a smaller set of clean (user approved) and larger set of noisy (unapproved) training patches of ROIs and background. The pre-trained deep-learning model is thereafter first trained on the large set of noisy patches, followed by refined training using the clean patches. Results: The framework is evaluated on fluorescence microscopy images from a large-scale drug screening experiment, brightfield images of immunohistochemistry-stained patient tissue samples, and malaria-infected human blood smears, as well as transmission electron microscopy images of cell sections. Compared to state-of-the-art and manual/visual assessment, the results show similar performance with maximal flexibility and minimal a priori information and user interaction. Conclusions: SimSearch quickly adapts to different data sets, which demonstrates the potential to speed up many microscopy-based experiments based on a small amount of user interaction. Significance: SimSearch can help biologists quickly extract informative regions and perform analyses on large datasets helping increase the throughput in a microscopy experiment.
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| 10. |
- Gupta, Ankit, et al.
(författare)
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Towards Better Guided Attention and Human Knowledge Insertion in Deep Convolutional Neural Networks
- 2022
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Konferensbidrag (refereegranskat)abstract
- Attention Branch Networks (ABNs) have been shown to simultaneously provide visual explanation and improve the performance of deep convolutional neural networks (CNNs). In this work, we introduce Multi-Scale Attention Branch Networks (MSABN), which enhance the resolution of the generated attention maps, and improve the performance. We evaluate MSABN on benchmark image recognition and fine-grained recognition datasets where we observe MSABN outperforms ABN and baseline models. We also introduce a new data augmentation strategy utilizing the attention maps to incorporate human knowledge in the form of bounding box annotations of the objects of interest. We show that even with a limited number of edited samples, a significant performance gain can be achieved with this strategy.
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