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    Applications of high-throughput clonogenic survival assays in high-LET particle microbeams.

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    Authors
    Georgantzoglou, A
    Merchant, Michael J
    Jeynes, J
    Mayhead, N
    Punia, N
    Butler, R
    Jena, R
    Affiliation
    Department of Oncology, Addenbrooke's Hospital, University of Cambridge , Cambridge , UK
    Issue Date
    2015
    
    Metadata
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    Abstract
    Charged particle therapy is increasingly becoming a valuable tool in cancer treatment, mainly due to the favorable interaction of particle radiation with matter. Its application is still limited due, in part, to lack of data regarding the radiosensitivity of certain cell lines to this radiation type, especially to high-linear energy transfer (LET) particles. From the earliest days of radiation biology, the clonogenic survival assay has been used to provide radiation response data. This method produces reliable data but it is not optimized for high-throughput microbeam studies with high-LET radiation where high levels of cell killing lead to a very low probability of maintaining cells' clonogenic potential. A new method, therefore, is proposed in this paper, which could potentially allow these experiments to be conducted in a high-throughput fashion. Cells are seeded in special polypropylene dishes and bright-field illumination provides cell visualization. Digital images are obtained and cell detection is applied based on corner detection, generating individual cell targets as x-y points. These points in the dish are then irradiated individually by a micron field size high-LET microbeam. Post-irradiation, time-lapse imaging follows cells' response. All irradiated cells are tracked by linking trajectories in all time-frames, based on finding their nearest position. Cell divisions are detected based on cell appearance and individual cell temporary corner density. The number of divisions anticipated is low due to the high probability of cell killing from high-LET irradiation. Survival curves are produced based on cell's capacity to divide at least four to five times. The process is repeated for a range of doses of radiation. Validation shows the efficiency of the proposed cell detection and tracking method in finding cell divisions.
    Citation
    Applications of high-throughput clonogenic survival assays in high-LET particle microbeams. 2015, 5:305 Front Oncol
    Journal
    Frontiers in Oncology
    URI
    http://hdl.handle.net/10541/596638
    DOI
    10.3389/fonc.2015.00305
    PubMed ID
    26835414
    Type
    Article
    Language
    en
    ISSN
    2234-943X
    ae974a485f413a2113503eed53cd6c53
    10.3389/fonc.2015.00305
    Scopus Count
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