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Automated microbeam observation environment for biological analysis-Custom portable environmental control applied to a vertical microbeam system.

ABSTRACT: Vertical Microbeams (VMB) are used to irradiate individual cells with low MeV energy ions. The irradiation of cells using VMBs requires cells to be removed from an incubator; this can cause physiological changes to cells because of the lower CO2 concentration, temperature and relative humidity outside of the incubator. Consequently, for experiments where cells require irradiation and observation for extended time periods, it is important to provide a controlled environment. The highly customised nature of the microscopes used on VMB systems means that there are no commercially available environmentally controlled microscope systems for VMB systems. The Automated Microbeam Observation Environment for Biological Analysis (AMOEBA) is a highly flexible modular environmental control system used to create incubator conditions on the end of a VMB. The AMOEBA takes advantage of the recent "maker" movement to create an open source control system that can be easily configured by the user to fit their control needs even beyond VMB applications. When applied to the task of controlling cell medium temperature, CO2 concentration and relative humidity on VMBs it creates a stable environment that allows cells to multiply on the end of a VMB over a period of 36 h, providing a low-cost (costing less than $2700 to build), customisable alternative to commercial time-lapse microscopy systems. AMOEBA adds the potential of VMBs to explore the long-term effects of radiation on single cells opening up new research areas for VMBs.

SUBMITTER: England MJ

PROVIDER: S-EPMC5836785 | biostudies-literature | 2017 Feb

REPOSITORIES: biostudies-literature

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Automated microbeam observation environment for biological analysis-Custom portable environmental control applied to a vertical microbeam system.

England Matthew J MJ Bigelow Alan W AW Merchant Michael J MJ Velliou Eirini E Welch David D Brenner David J DJ Kirkby Karen J KJ

Sensors and actuators. B, Chemical 20160816

Vertical Microbeams (VMB) are used to irradiate individual cells with low MeV energy ions. The irradiation of cells using VMBs requires cells to be removed from an incubator; this can cause physiological changes to cells because of the lower CO<sub>2</sub> concentration, temperature and relative humidity outside of the incubator. Consequently, for experiments where cells require irradiation and observation for extended time periods, it is important to provide a controlled environment. The highly ...[more]

PMID: 29515291

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Project description:AimsEnhanced Biological Phosphorus Removal (EBPR) is a technology widely used in wastewater treatment to remove phosphorus (P) and prevent eutrophication. Establishing its operating efficiency and stability is an active research field that has generated almost 3000 publications in the last 40 years. Due to its size, including over 119 review articles, it is an example of a field where it becomes increasingly difficult to manually recognize its key research contributions, especially for non-experts or newcomers. Therefore, this work included two distinct but complementary objectives. First, to assemble for the first time a collection of bibliometric techniques into a framework for automating the article selection process when preparing a literature review (section 2). Second, to demonstrate it by applying it to the field of EBPR, producing a bibliometric analysis and a review of the key findings of EBPR research over time (section 3).FindingsThe joint analysis of citation networks, keywords, citation profiles, as well as of specific benchmarks for the identification of highly-cited publications revealed 12 research topics. Their content and evolution could be manually reviewed using a selection of articles consisting of approximately only 5% of the original set of publications. The largest topics addressed the identification of relevant microorganisms, the characterization of their metabolism, including denitrification and the competition between them (Clusters A-D). Emerging and influential topics, as determined by different citation indicators and temporal analysis, were related to volatile fatty acid production, P-recovery from waste activated sludge and aerobic granules for better process efficiency and stability (Clusters F-H).ConclusionsThe framework enabled key contributions in each of the constituent topics to be highlighted in a way that may have otherwise been biased by conventional citation-based ranking. Further, it reduced the need for manual input and a priori expertise compared to a traditional literature review. Hence, in an era of accelerated production of information and publications, this work contributed to the way that we are able to use computer-aided approaches to curate information and manage knowledge.

Project description:Exposure to cigarette smoke is a leading cause of lung diseases including chronic obstructive pulmonary disease and cancer. Cigarette smoke is a complex aerosol containing over 6000 chemicals, and thus it is difficult to determine individual contributions to overall toxicity, and the molecular mechanisms by which smoke constituents exert their effects. We selected three well-known harmful and potentially harmful constituents (HPHCs) in tobacco smoke: acrolein, formaldehyde and catechol and established a High Content Screening (HCS) method using normal human bronchial epithelial cells, which are the first bronchial cells in contact with cigarette smoke. The impact of each HPHC was investigated using 13 multi-parametric indicators of cellular toxicity and complemented with a microarray-based whole transcriptome analysis followed by a computational approach leveraging mechanistic network models to identify and quantify perturbed molecular pathways. HPHCs were evaluated over a wide range of concentrations and at different exposure time points (4 h, 8 h, and 24 h). By High Content Screening, the toxic effects of the three HPHCs could only be observed at the highest doses. Whole genome transcriptomics unraveled toxicity mechanisms at lower doses and earlier time points. The most prevalent toxicity mechanisms observed were: DNA damage/growth arrest, oxidative stress, mitochondrial stress and apoptosis/necrosis. In summary, combination of multiple toxicological endpoints with a systems-based impact assessment allows for a more robust scientific basis for the toxicological assessment of HPHCs that allows insight into time- and dose-dependent molecular perturbations of specific biological pathways. This approach allowed us to establish an in vitro Systems Toxicology platform that can be applied to a broader selection of HPHCs and their mixtures and can serve more generally as the basis for testing the impact of other environmental toxicants on normal bronchial epithelial cells.

Dataset Information

Automated microbeam observation environment for biological analysis-Custom portable environmental control applied to a vertical microbeam system.

Publications

Automated microbeam observation environment for biological analysis-Custom portable environmental control applied to a vertical microbeam system.

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