Cellular tracking in time-lapse phase contrast images

The quantitative analysis of live cells is a key issue in evaluating biological processes. The current clinical practice involves the application of a tedious and time consuming manual tracking procedure on large amount of data. As a result, automatic tracking systems are currently developed and evaluated. However, problems caused by cellular division, agglomeration, Brownian motion and topology changes are difficult issues that have to be accommodated by automatic tracking techniques. In this paper, we detail the development of a fully automated multi-target tracking system that is able to deal with Brownian motion and cellular division. During the tracking process our approach includes the neighbourhood relationship and motion history to enforce the cellular tracking continuity in the spatial and temporal domain. The experimental results reported in this paper indicate that our method is able to accurately track cellular structures in time-lapse data

A novel framework for cellular tracking and mitosis detection in dense phase contrast microscopy images

The aim of this paper is to detail the development of a novel tracking framework that is able to extract the cell motility indicators and to determine the cellular division (mitosis) events in large time-lapse phase-contrast image sequences. To address the challenges induced by non-structured (random) motion, cellular agglomeration, and cellular mitosis, the process of automatic (unsupervised) cell tracking is carried out in a sequential manner, where the inter-frame cell association is achieved by assessing the variation in the local cellular structures in consecutive frames of the image sequence. In our study a strong emphasis has been placed on the robust use of the topological information in the cellular tracking process and in the development of targeted pattern recognition techniques that were designed to redress the problems caused by segmentation errors, and to precisely identify mitosis using a backward (reversed) tracking strategy. The proposed algorithm has been evaluated on dense phase contrast cellular data and the experimental results indicate that the proposed algorithm is able to accurately track epithelial and endothelial cells in time-lapse image sequences that are characterized by low contrast and high level of noise. Our algorithm achieved 86.10% overall tracking accuracy and 90.12% mitosis detection accuracy

Automatic cellular segmentation in time-lapse phase contrast images

The process of cellular detection and tracking is a key task in the analysis of cellular motility and proliferation. The current clinical procedure involves a time consuming procedure that requires the manual annotation of cells in sequences of time-lapse phase contrast microscopy images. With the development of modern imaging modalities, the amount of data to be interpreted by biologists is constantly increasing, thus the development of automatic techniques that are able to detect cellular structures in large image sequences is more necessary than ever before. Robust cellular detection represents the first step in the development of cellular tracking algorithms and one of the objectives of our work was focused on the development of an automatic technique that is able to segment the cells in various sequences of cellular data. The proposed segmentation framework adaptively determines the criteria to separate the cells and the background and additional morphological operations are applied to detect the initial structures that define the cells in each image of the sequence. The initial segmentation results are refined by applying motion consistency constraints to detect the cells that are missed by the initial segmentation process due to factors such as image noise and low contrast. In our experiments we have applied the proposed segmentation framework to NE4C, MDCK and HUVEC cellular data. A number of experimental results are illustrated in Figure 1

A novel framework for tracking in-vitro cells in time-lapse phase contrast data

With the proliferation of modern microscopy imaging technologies the amount of data that has to be analysed by biologists is constantly increasing and as a result the development of automatic approaches that are able to track cellular structures in timelapse images has become an important field of research. The aim of this paper is to detail the development of a novel tracking framework that is designed to extract the cell motility indicators in phase-contrast image sequences. To address issues that are caused by nonstructured (random) motion and cellular agglomeration, cell tracking is formulated as a sequential process where the inter-frame cell association is achieved by assessing the variation in the local structures contained in consecutive frames of the image sequence. We have evaluated the proposed algorithm on dense phase contrast cellular data and the reported results indicate that the developed algorithm is able to accurately track MadinDarby Canine Kidney (MDCK) Epithelial Cells in image data that is characterised by low contrast and high level of noise

Determining cellular CTCF and cohesin abundances to constrain 3D genome models.

Achieving a quantitative and predictive understanding of 3D genome architecture remains a major challenge, as it requires quantitative measurements of the key proteins involved. Here, we report the quantification of CTCF and cohesin, two causal regulators of topologically associating domains (TADs) in mammalian cells. Extending our previous imaging studies (Hansen et al., 2017), we estimate bounds on the density of putatively DNA loop-extruding cohesin complexes and CTCF binding site occupancy. Furthermore, co-immunoprecipitation studies of an endogenously tagged subunit (Rad21) suggest the presence of cohesin dimers and/or oligomers. Finally, based on our cell lines with accurately measured protein abundances, we report a method to conveniently determine the number of molecules of any Halo-tagged protein in the cell. We anticipate that our results and the established tool for measuring cellular protein abundances will advance a more quantitative understanding of 3D genome organization, and facilitate protein quantification, key to comprehend diverse biological processes

A quantitative map of nuclear pore assembly reveals two distinct mechanisms

Understanding how the nuclear pore complex (NPC) assembles is of fundamental importance to grasp the mechanisms behind its essential function and understand its role during evolution of eukaryotes1–4. While we know that at least two NPC assembly pathways exist, one during exit from mitosis and one during nuclear growth in interphase, we currently lack a quantitative map of their molecular events. Here, we use fluorescence correlation spectroscopy (FCS) calibrated live imaging of endogenously fluorescently-tagged nucleoporins to map the changes in composition and stoichiometry of seven major modules of the human NPC during its assembly in single dividing cells. This systematic quantitative map reveals that the two assembly pathways employ strikingly different molecular mechanisms, inverting the order of addition of two large structural components, the central ring complex and nuclear filaments. Our dynamic stoichiometry data allows us to perform the first computational simulation that predicts the structure of postmitotic NPC assembly intermediates

Socializing One Health: an innovative strategy to investigate social and behavioral risks of emerging viral threats

In an effort to strengthen global capacity to prevent, detect, and control infectious diseases in animals and people, the United States Agency for International Development’s (USAID) Emerging Pandemic Threats (EPT) PREDICT project funded development of regional, national, and local One Health capacities for early disease detection, rapid response, disease control, and risk reduction. From the outset, the EPT approach was inclusive of social science research methods designed to understand the contexts and behaviors of communities living and working at human-animal-environment interfaces considered high-risk for virus emergence. Using qualitative and quantitative approaches, PREDICT behavioral research aimed to identify and assess a range of socio-cultural behaviors that could be influential in zoonotic disease emergence, amplification, and transmission. This broad approach to behavioral risk characterization enabled us to identify and characterize human activities that could be linked to the transmission dynamics of new and emerging viruses. This paper provides a discussion of implementation of a social science approach within a zoonotic surveillance framework. We conducted in-depth ethnographic interviews and focus groups to better understand the individual- and community-level knowledge, attitudes, and practices that potentially put participants at risk for zoonotic disease transmission from the animals they live and work with, across 6 interface domains. When we asked highly-exposed individuals (ie. bushmeat hunters, wildlife or guano farmers) about the risk they perceived in their occupational activities, most did not perceive it to be risky, whether because it was normalized by years (or generations) of doing such an activity, or due to lack of information about potential risks. Integrating the social sciences allows investigations of the specific human activities that are hypothesized to drive disease emergence, amplification, and transmission, in order to better substantiate behavioral disease drivers, along with the social dimensions of infection and transmission dynamics. Understanding these dynamics is critical to achieving health security--the protection from threats to health-- which requires investments in both collective and individual health security. Involving behavioral sciences into zoonotic disease surveillance allowed us to push toward fuller community integration and engagement and toward dialogue and implementation of recommendations for disease prevention and improved health security

An adaptive motion segmentation for automated video surveillance

This paper presents an adaptive motion segmentation algorithm utilizing spatiotemporal information of three most recent frames. The algorithm initially extracts the moving edges applying a novel flexible edge matching technique which makes use of a combined distance transformation image. Then watershed-based iterative algorithm is employed to segment the moving object region from the extracted moving edges. The challenges of existing three-frame-based methods include slow movement, edge localization error, minor movement of camera, and homogeneity of background and foreground region. The proposed method represents edges as segments and uses a flexible edge matching algorithm to deal with edge localization error and minor movement of camera. The combined distance transformation image works in favor of accumulating gradient information of overlapping region which effectively improves the sensitivity to slow movement. The segmentation algorithm uses watershed, gradient information of difference image, and extracted moving edges. It helps to segment moving object region with more accurate boundary even some part of the moving edges cannot be detected due to region homogeneity or other reasons during the detection step. Experimental results using different types of video sequences are presented to demonstrate the efficiency and accuracy of the proposed method.</p

Segmentation of moving object for content based applications

This paper presents an edge segment based moving object segmentation algorithm independent of background model. The proposed method detects moving edges using three most recent frames where moving regions are extracted by employing a watershed based algorithm. Extracted moving segments help to ensure good visual quality even in limited bit rate multimedia communications by incorporating priority basedtransmission.</p