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  1. Article ; Online: A Review of Super-Resolution Single-Molecule Localization Microscopy Cluster Analysis and Quantification Methods.

    Khater, Ismail M / Nabi, Ivan Robert / Hamarneh, Ghassan

    Patterns (New York, N.Y.)

    2020  Volume 1, Issue 3, Page(s) 100038

    Abstract: Single-molecule localization microscopy (SMLM) is a relatively new imaging modality, winning the 2014 Nobel Prize in Chemistry, and considered as one of the key super-resolution techniques. SMLM resolution goes beyond the diffraction limit of light ... ...

    Abstract Single-molecule localization microscopy (SMLM) is a relatively new imaging modality, winning the 2014 Nobel Prize in Chemistry, and considered as one of the key super-resolution techniques. SMLM resolution goes beyond the diffraction limit of light microscopy and achieves resolution on the order of 10-20 nm. SMLM thus enables imaging single molecules and study of the low-level molecular interactions at the subcellular level. In contrast to standard microscopy imaging that produces 2D pixel or 3D voxel grid data, SMLM generates big data of 2D or 3D point clouds with millions of localizations and associated uncertainties. This unprecedented breakthrough in imaging helps researchers employ SMLM in many fields within biology and medicine, such as studying cancerous cells and cell-mediated immunity and accelerating drug discovery. However, SMLM data quantification and interpretation methods have yet to keep pace with the rapid advancement of SMLM imaging. Researchers have been actively exploring new computational methods for SMLM data analysis to extract biosignatures of various biological structures and functions. In this survey, we describe the state-of-the-art clustering methods adopted to analyze and quantify SMLM data and examine the capabilities and shortcomings of the surveyed methods. We classify the methods according to (1) the biological application (i.e., the imaged molecules/structures), (2) the data acquisition (such as imaging modality, dimension, resolution, and number of localizations), and (3) the analysis details (2D versus 3D, field of view versus region of interest, use of machine-learning and multi-scale analysis, biosignature extraction, etc.). We observe that the majority of methods that are based on second-order statistics are sensitive to noise and imaging artifacts, have not been applied to 3D data, do not leverage machine-learning formulations, and are not scalable for big-data analysis. Finally, we summarize state-of-the-art methodology, discuss some key open challenges, and identify future opportunities for better modeling and design of an integrated computational pipeline to address the key challenges.
    Language English
    Publishing date 2020-06-12
    Publishing country United States
    Document type Journal Article ; Review
    ISSN 2666-3899
    ISSN (online) 2666-3899
    DOI 10.1016/j.patter.2020.100038
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Book ; Online: AI-based analysis of super-resolution microscopy

    Nabi, Ivan R. / Cardoen, Ben / Khater, Ismail M. / Gao, Guang / Wong, Timothy H. / Hamarneh, Ghassan

    Biological discovery in the absence of ground truth

    2023  

    Abstract: The nanoscale resolution of super-resolution microscopy has now enabled the use of fluorescent based molecular localization tools to study whole cell structural biology. Machine learning based analysis of super-resolution data offers tremendous potential ...

    Abstract The nanoscale resolution of super-resolution microscopy has now enabled the use of fluorescent based molecular localization tools to study whole cell structural biology. Machine learning based analysis of super-resolution data offers tremendous potential for discovery of new biology, that by definition is not known and lacks ground truth. Herein, we describe the application of weakly supervised learning paradigms to super-resolution microscopy and its potential to enable the accelerated exploration of the molecular architecture of subcellular macromolecules and organelles.

    Comment: 14 pages, 3 figures
    Keywords Quantitative Biology - Subcellular Processes ; Computer Science - Artificial Intelligence ; Computer Science - Computer Vision and Pattern Recognition ; Computer Science - Machine Learning ; Physics - Biological Physics ; Quantitative Biology - Quantitative Methods
    Publishing date 2023-05-26
    Publishing country us
    Document type Book ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article ; Online: Single molecule network analysis identifies structural changes to caveolae and scaffolds due to mutation of the caveolin-1 scaffolding domain.

    Wong, Timothy H / Khater, Ismail M / Joshi, Bharat / Shahsavari, Mona / Hamarneh, Ghassan / Nabi, Ivan R

    Scientific reports

    2021  Volume 11, Issue 1, Page(s) 7810

    Abstract: Caveolin-1 (CAV1), the caveolae coat protein, also associates with non-caveolar scaffold domains. Single molecule localization microscopy (SMLM) network analysis distinguishes caveolae and three scaffold domains, hemispherical S2 scaffolds and smaller ... ...

    Abstract Caveolin-1 (CAV1), the caveolae coat protein, also associates with non-caveolar scaffold domains. Single molecule localization microscopy (SMLM) network analysis distinguishes caveolae and three scaffold domains, hemispherical S2 scaffolds and smaller S1B and S1A scaffolds. The caveolin scaffolding domain (CSD) is a highly conserved hydrophobic region that mediates interaction of CAV1 with multiple effector molecules. F92A/V94A mutation disrupts CSD function, however the structural impact of CSD mutation on caveolae or scaffolds remains unknown. Here, SMLM network analysis quantitatively shows that expression of the CAV1 CSD F92A/V94A mutant in CRISPR/Cas CAV1 knockout MDA-MB-231 breast cancer cells reduces the size and volume and enhances the elongation of caveolae and scaffold domains, with more pronounced effects on S2 and S1B scaffolds. Convex hull analysis of the outer surface of the CAV1 point clouds confirms the size reduction of CSD mutant CAV1 blobs and shows that CSD mutation reduces volume variation amongst S2 and S1B CAV1 blobs at increasing shrink values, that may reflect retraction of the CAV1 N-terminus towards the membrane, potentially preventing accessibility of the CSD. Detection of point mutation-induced changes to CAV1 domains highlights the utility of SMLM network analysis for mesoscale structural analysis of oligomers in their native environment.
    MeSH term(s) Caveolin 1/chemistry ; Cell Line ; Humans ; Mutation ; Protein Conformation ; Protein Domains/genetics
    Chemical Substances CAV1 protein, human ; Caveolin 1
    Language English
    Publishing date 2021-04-08
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-021-86770-6
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Identification of caveolin-1 domain signatures via machine learning and graphlet analysis of single-molecule super-resolution data.

    Khater, Ismail M / Meng, Fanrui / Nabi, Ivan Robert / Hamarneh, Ghassan

    Bioinformatics (Oxford, England)

    2019  Volume 35, Issue 18, Page(s) 3468–3475

    Abstract: Motivation: Network analysis and unsupervised machine learning processing of single-molecule localization microscopy of caveolin-1 (Cav1) antibody labeling of prostate cancer cells identified biosignatures and structures for caveolae and three distinct ... ...

    Abstract Motivation: Network analysis and unsupervised machine learning processing of single-molecule localization microscopy of caveolin-1 (Cav1) antibody labeling of prostate cancer cells identified biosignatures and structures for caveolae and three distinct non-caveolar scaffolds (S1A, S1B and S2). To obtain further insight into low-level molecular interactions within these different structural domains, we now introduce graphlet decomposition over a range of proximity thresholds and show that frequency of different subgraph (k = 4 nodes) patterns for machine learning approaches (classification, identification, automatic labeling, etc.) effectively distinguishes caveolae and scaffold blobs.
    Results: Caveolae formation requires both Cav1 and the adaptor protein CAVIN1 (also called PTRF). As a supervised learning approach, we applied a wide-field CAVIN1/PTRF mask to CAVIN1/PTRF-transfected PC3 prostate cancer cells and used the random forest classifier to classify blobs based on graphlet frequency distribution (GFD). GFD of CAVIN1/PTRF-positive (PTRF+) and -negative Cav1 clusters showed poor classification accuracy that was significantly improved by stratifying the PTRF+ clusters by either number of localizations or volume. Low classification accuracy (<50%) of large PTRF+ clusters and caveolae blobs identified by unsupervised learning suggests that their GFD is specific to caveolae. High classification accuracy for small PTRF+ clusters and caveolae blobs argues that CAVIN1/PTRF associates not only with caveolae but also non-caveolar scaffolds. At low proximity thresholds (50-100 nm), the caveolae groups showed reduced frequency of highly connected graphlets and increased frequency of completely disconnected graphlets. GFD analysis of single-molecule localization microscopy Cav1 clusters defines changes in structural organization in caveolae and scaffolds independent of association with CAVIN1/PTRF.
    Supplementary information: Supplementary data are available at Bioinformatics online.
    MeSH term(s) Caveolae ; Caveolin 1 ; Humans ; Machine Learning ; Male ; Prostatic Neoplasms ; RNA-Binding Proteins
    Chemical Substances Caveolin 1 ; RNA-Binding Proteins
    Language English
    Publishing date 2019-03-13
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 1422668-6
    ISSN 1367-4811 ; 1367-4803
    ISSN (online) 1367-4811
    ISSN 1367-4803
    DOI 10.1093/bioinformatics/btz113
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Caveolae and scaffold detection from single molecule localization microscopy data using deep learning.

    Khater, Ismail M / Aroca-Ouellette, Stephane T / Meng, Fanrui / Nabi, Ivan Robert / Hamarneh, Ghassan

    PloS one

    2019  Volume 14, Issue 8, Page(s) e0211659

    Abstract: Caveolae are plasma membrane invaginations whose formation requires caveolin-1 (Cav1), the adaptor protein polymerase I, and the transcript release factor (PTRF or CAVIN1). Caveolae have an important role in cell functioning, signaling, and disease. In ... ...

    Abstract Caveolae are plasma membrane invaginations whose formation requires caveolin-1 (Cav1), the adaptor protein polymerase I, and the transcript release factor (PTRF or CAVIN1). Caveolae have an important role in cell functioning, signaling, and disease. In the absence of CAVIN1/PTRF, Cav1 forms non-caveolar membrane domains called scaffolds. In this work, we train machine learning models to automatically distinguish between caveolae and scaffolds from single molecule localization microscopy (SMLM) data. We apply machine learning algorithms to discriminate biological structures from SMLM data. Our work is the first that is leveraging machine learning approaches (including deep learning models) to automatically identifying biological structures from SMLM data. In particular, we develop and compare three binary classification methods to identify whether or not a given 3D cluster of Cav1 proteins is a caveolae. The first uses a random forest classifier applied to 28 hand-crafted/designed features, the second uses a convolutional neural net (CNN) applied to a projection of the point clouds onto three planes, and the third uses a PointNet model, a recent development that can directly take point clouds as its input. We validate our methods on a dataset of super-resolution microscopy images of PC3 prostate cancer cells labeled for Cav1. Specifically, we have images from two cell populations: 10 PC3 and 10 CAVIN1/PTRF-transfected PC3 cells (PC3-PTRF cells) that form caveolae. We obtained a balanced set of 1714 different cellular structures. Our results show that both the random forest on hand-designed features and the deep learning approach achieve high accuracy in distinguishing the intrinsic features of the caveolae and non-caveolae biological structures. More specifically, both random forest and deep CNN classifiers achieve classification accuracy reaching 94% on our test set, while the PointNet model only reached 83% accuracy. We also discuss the pros and cons of the different approaches.
    MeSH term(s) Caveolae/metabolism ; Deep Learning ; Humans ; Image Processing, Computer-Assisted/methods ; PC-3 Cells ; Single Molecule Imaging
    Language English
    Publishing date 2019-08-26
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1932-6203
    ISSN (online) 1932-6203
    DOI 10.1371/journal.pone.0211659
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Super-resolution modularity analysis shows polyhedral caveolin-1 oligomers combine to form scaffolds and caveolae.

    Khater, Ismail M / Liu, Qian / Chou, Keng C / Hamarneh, Ghassan / Nabi, Ivan Robert

    Scientific reports

    2019  Volume 9, Issue 1, Page(s) 9888

    Abstract: Caveolin-1 (Cav1), the coat protein for caveolae, also forms non-caveolar Cav1 scaffolds. Single molecule Cav1 super-resolution microscopy analysis previously identified caveolae and three distinct scaffold domains: smaller S1A and S2B scaffolds and ... ...

    Abstract Caveolin-1 (Cav1), the coat protein for caveolae, also forms non-caveolar Cav1 scaffolds. Single molecule Cav1 super-resolution microscopy analysis previously identified caveolae and three distinct scaffold domains: smaller S1A and S2B scaffolds and larger hemispherical S2 scaffolds. Application here of network modularity analysis of SMLM data for endogenous Cav1 labeling in HeLa cells shows that small scaffolds combine to form larger scaffolds and caveolae. We find modules within Cav1 blobs by maximizing the intra-connectivity between Cav1 molecules within a module and minimizing the inter-connectivity between Cav1 molecules across modules, which is achieved via spectral decomposition of the localizations adjacency matrix. Features of modules are then matched with intact blobs to find the similarity between the module-blob pairs of group centers. Our results show that smaller S1A and S1B scaffolds are made up of small polygons, that S1B scaffolds correspond to S1A scaffold dimers and that caveolae and hemispherical S2 scaffolds are complex, modular structures formed from S1B and S1A scaffolds, respectively. Polyhedral interactions of Cav1 oligomers, therefore, leads progressively to the formation of larger and more complex scaffold domains and the biogenesis of caveolae.
    MeSH term(s) Caveolae/metabolism ; Caveolin 1/metabolism ; Cell Line, Tumor ; Cell Membrane/metabolism ; HeLa Cells ; Humans ; Microscopy/methods ; Single Molecule Imaging/methods
    Chemical Substances CAV1 protein, human ; Caveolin 1
    Language English
    Publishing date 2019-07-08
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-019-46174-z
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Super Resolution Network Analysis Defines the Molecular Architecture of Caveolae and Caveolin-1 Scaffolds.

    Khater, Ismail M / Meng, Fanrui / Wong, Timothy H / Nabi, Ivan Robert / Hamarneh, Ghassan

    Scientific reports

    2018  Volume 8, Issue 1, Page(s) 9009

    Abstract: Quantitative approaches to analyze the large data sets generated by single molecule localization super-resolution microscopy (SMLM) are limited. We developed a computational pipeline and applied it to analyzing 3D point clouds of SMLM localizations ( ... ...

    Abstract Quantitative approaches to analyze the large data sets generated by single molecule localization super-resolution microscopy (SMLM) are limited. We developed a computational pipeline and applied it to analyzing 3D point clouds of SMLM localizations (event lists) of the caveolar coat protein, caveolin-1 (Cav1), in prostate cancer cells differentially expressing CAVIN1 (also known as PTRF), that is also required for caveolae formation. High degree (strongly-interacting) points were removed by an iterative blink merging algorithm and Cav1 network properties were compared with randomly generated networks to retain a sub-network of geometric structures (or blobs). Machine-learning based classification extracted 28 quantitative features describing the size, shape, topology and network characteristics of ∼80,000 blobs. Unsupervised clustering identified small S1A scaffolds corresponding to SDS-resistant Cav1 oligomers, as yet undescribed larger hemi-spherical S2 scaffolds and, only in CAVIN1-expressing cells, spherical, hollow caveolae. Multi-threshold modularity analysis suggests that S1A scaffolds interact to form larger scaffolds and that S1A dimers group together, in the presence of CAVIN1, to form the caveolae coat.
    MeSH term(s) Algorithms ; Caveolae/metabolism ; Caveolin 1/metabolism ; Humans ; Image Processing, Computer-Assisted/instrumentation ; Image Processing, Computer-Assisted/methods ; Imaging, Three-Dimensional/instrumentation ; Imaging, Three-Dimensional/methods ; Male ; Microscopy/instrumentation ; Microscopy/methods ; PC-3 Cells ; Prostatic Neoplasms/genetics ; Prostatic Neoplasms/metabolism ; Prostatic Neoplasms/pathology ; Protein Binding ; Protein Interaction Maps ; RNA-Binding Proteins/genetics ; RNA-Binding Proteins/metabolism ; Software
    Chemical Substances CAVIN1 protein, human ; Caveolin 1 ; RNA-Binding Proteins
    Language English
    Publishing date 2018-06-13
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-018-27216-4
    Database MEDical Literature Analysis and Retrieval System OnLINE

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