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Detecting sequential bond formation using three-dimensional thermal fluctuation analysis

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    SYSNO ASEP0333759
    Document TypeJ - Journal Article
    R&D Document TypeJournal Article
    Subsidiary JČlánek ve WOS
    TitleDetecting sequential bond formation using three-dimensional thermal fluctuation analysis
    Author(s) Bartsch, T. (US)
    Fisinger, S. (US)
    Kochanczyk, M.D. (US)
    Huang, R. (US)
    Jonáš, Alexandr (UPT-D) RID, SAI, ORCID
    Florin, E. L. (DE)
    Source TitleChemPhysChem. - : Wiley - ISSN 1439-4235
    Roč. 10, 9-10 (2009), s. 1541-1547
    Number of pages7 s.
    Languageeng - English
    CountryDE - Germany
    Keywordsbiophysics ; mechanical properties ; optical tweezers ; single-molecule studies ; thermal fluctuations
    Subject RIVBO - Biophysics
    CEZAV0Z20650511 - UPT-D (2005-2011)
    UT WOS000267928100028
    DOI10.1002/cphc.200900211
    AnnotationWe present a novel experimental method that solves two key problems in nondestructive mechanical studies of small biomolecules at the single-molecule level, namely the confirmation of single-molecule conditions and the discrimination against nonspecific binding. A biotin avidin ligand-receptor couple is spanned between a glass slide and a 1 mu m latex particles using short linker molecules. Optical tweezers are used to initiate bond formation and to follow the particles thermal position fluctuations with nanometer spatial and microsecond temporal resolution. Here we show that each step in the specific binding process leads to an abrupt change in the magnitude of the particle's thermal position fluctuations, allowing us to count the number of bonds formed one by one. Moreover, three-dimensional position histograms calculated from the particle's fluctuations can be separated into well-defined categories reflecting different binding conditions.
    WorkplaceInstitute of Scientific Instruments
    ContactMartina Šillerová, sillerova@ISIBrno.Cz, Tel.: 541 514 178
    Year of Publishing2010
Number of the records: 1  

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