The identification of cell borders (‘segmentation’) in microscopy images constitutes a bottleneck for large-scale experiments. For the model organism Saccharomyces cerevisiae, current segmentation methods face challenges when cells bud, crowd, or exhibit irregular features. We present a convolutional neural network (CNN) named YeaZ, the underlying training set of high-quality segmented yeast images (>10 000 cells) including mutants, stressed cells, and time courses, as well as a graphical user interface and a web application (www.quantsysbio.com/data-and-software) to efficiently employ, test, and expand the system. A key feature is a cell-cell boundary test which avoids the need for fluorescent markers. Our CNN is highly accurate, including for buds, and outperforms existing methods on benchmark images, indicating it transfers well to other conditions. To demonstrate how efficient large-scale image processing uncovers new biology, we analyze the geometries of ≈2200 wild-type and cyclin mutant cells and find that morphogenesis control occurs unexpectedly early and gradually.
The ability to independently control the expression of different genes is important for quantitative biology. Using budding yeast, we characterize GAL1pr, GALL, MET3pr, CUP1pr, PHO5pr, tetOpr, terminator-tetOpr, Z3EV, blue-light inducible optogenetic systems El222-LIP, El222-GLIP, and red-light inducible PhyB-PIF3. We report kinetic parameters, noise scaling, impact on growth, and the fundamental leakiness of each system using an intuitive unit, maxGAL1. We uncover disadvantages of widely used tools, e.g., nonmonotonic activity of MET3pr and GALL, slow off kinetics of the doxycycline- and estradiol-inducible systems tetOpr and Z3EV, and high variability of PHO5pr and red-light activated PhyB-PIF3 system. We introduce two previously uncharacterized systems: strongLOV, a more light-sensitive El222 mutant, and ARG3pr, which is induced in the absence of arginine or presence of methionine. To demonstrate fine control over gene circuits, we experimentally tune the time between cell cycle Start and mitosis, artificially simulating near-wild-type timing. All strains, constructs, code, and data (https://promoter-benchmark.epfl.ch/) are made available.
Virtual Private Networks (VPNs) are increasingly used to build logically isolated networks. However, existing VPN designs and deployments neglect the problem of traffic analysis and covert channels. Hence, there are many ways to infer information from VPN traffic without decrypting it. Many proposals were made to mitigate network covert channels, but previous works remained largely theoretical or resulted in prohibitively high padding overhead and performance penalties. In this work, we (1) analyse the impact of covert channels in IPsec, (2) present several improved and novel approaches for covert channel mitigation in IPsec, (3) propose and implement a system for dynamic performance trade-offs, and (4) implement our design in the Linux IPsec stack and evaluate its performance for different types of traffic and mitigation policies. At only 24% overhead, our prototype enforces tight informationtheoretic bounds on all information leakage.
words)The processing of microscopy images constitutes a bottleneck for large-scale experiments. A critical step is the establishment of cell borders ('segmentation'), which is required for a range of applications such as growth or fluorescent reporter measurements. For the model organism budding yeast (Saccharomyces cerevisiae), a number of methods for segmentation exist. However, in experiments involving multiple cell cycles, stress, or various mutants, cells crowd or exhibit irregular visible features, which necessitate frequent manual corrections. Furthermore, budding events are visually subtle but important to detect. Convolutional neural networks (CNNs) have been successfully employed for a range of image processing applications. They require large, diverse training sets. Here, we present i) the first set of publicly available, high-quality segmented yeast images (>10'000 cells) including mutants, stressed cells, and time courses, ii) a corresponding U-Net-based CNN, iii) a Python-based graphical user interface (GUI) to efficiently use the system, and iv) a web application to test it (www.quantsysbio.com). A key feature is a cell-cell boundary test which avoids the need for additional input from fluorescent channels. A bipartite graph matching algorithm tracks cells in time with high reliability. Our network is highly accurate and outperforms existing methods on benchmark images recorded by others, suggesting it transfers well to other conditions. Furthermore, new buds are detected early with high reliability. We apply the system to detect differences in geometry between wild-type and cyclin mutant cells. Our results indicate that morphogenesis control occurs unexpectedly early in the cell cycle and is gradual, demonstrating how the efficient processing of large numbers of cells uncovers new biology. Our system can serve as a resource to the community, expanded continuously with new images. Furthermore, the techniques we develop here are likely to be useful for other organisms as well.2 Dietler et al.YeaZ: A CNN for highly accurate, label-free segmentation of yeast images Abstract (150 words)The identification of cell borders ('segmentation') in microscopy images constitutes a bottleneck for large-scale experiments. For the model organism Saccharomyces cerevisiae, current segmentation methods face challenges when cells bud, crowd, or exhibit irregular features. Here, we present i) the first set of publicly available, high-quality segmented yeast images (>10'000 cells) including mutants, stressed cells, and time courses, ii) a corresponding convolutional neural network (CNN), iii) a graphical user interface and a web application (www.quantsysbio.com) to efficiently employ, test, and expand the system. A key feature is a cell-cell boundary test which avoids the need for fluorescent markers. Our CNN is highly accurate, including for buds, and outperforms existing methods on benchmark images, indicating it transfers well to other conditions. To demonstrate how efficient, large-scale image processing uncove...
Trusted Virtual Domains (TVDs) provide a secure IT infrastructure offering a homogeneous and transparent enforcement of access control policies on data and network resources. In this article, we give an overview of the fundamental ideas and basic concepts behind TVDs, present a realization of TVDs, and discuss application scenarios
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.