Dynamic imaging of single biomolecular interaction using flow control and TIRFM

T. Arakawa, T. Sameshima, Y. Sato, Y. Sumiyoshi, T. Ueno, Y. Shirasaki, T. Funatsu, Shuichi Shoji

    Research output: Chapter in Book/Report/Conference proceedingConference contribution

    Abstract

    A rapid multi-reagents switching microvalve system integrated with a total internal reflection fluorescence microscopy (TIRFM) was developed for real time imaging of a single protein behavior. The binding and dissociation process between a chaperonin GroEL and cochaperonin GroES was observed in this TIRFM microfluidic system. For single molecular imaging in this system, biotinylated GroEL (D490C) was immobilized to the glass microchannel surface through streptavidin and biotinylated BSA. A solution including 1 nM IC5-GroES was introduced for 100 ms into the observation area and then washed out with 2 mM ATP buffer solution. Fluorescence spots of IC5-GroES appeared after rapid solution switching, and disappeared several seconds later. As a result, we succeeded in detecting fluorescence signal from single molecules in TIRFM microfluidic system.

    Original languageEnglish
    Title of host publication12th International Conference on Miniaturized Systems for Chemistry and Life Sciences - The Proceedings of MicroTAS 2008 Conference
    PublisherChemical and Biological Microsystems Society
    Pages296-298
    Number of pages3
    Publication statusPublished - 2008
    Event12th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2008 - San Diego, CA
    Duration: 2008 Oct 122008 Oct 16

    Other

    Other12th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2008
    CitySan Diego, CA
    Period08/10/1208/10/16

    Fingerprint

    Fluorescence microscopy
    Flow control
    Imaging techniques
    Microfluidics
    Fluorescence
    Chaperonins
    Molecular imaging
    Streptavidin
    Switching systems
    Adenosinetriphosphate
    Microchannels
    Buffers
    Adenosine Triphosphate
    Proteins
    Glass
    Molecules

    Keywords

    • Microfluidic device
    • Single biomolecular imaging
    • TIRFM

    ASJC Scopus subject areas

    • Chemical Engineering (miscellaneous)
    • Bioengineering

    Cite this

    Arakawa, T., Sameshima, T., Sato, Y., Sumiyoshi, Y., Ueno, T., Shirasaki, Y., ... Shoji, S. (2008). Dynamic imaging of single biomolecular interaction using flow control and TIRFM. In 12th International Conference on Miniaturized Systems for Chemistry and Life Sciences - The Proceedings of MicroTAS 2008 Conference (pp. 296-298). Chemical and Biological Microsystems Society.

    Dynamic imaging of single biomolecular interaction using flow control and TIRFM. / Arakawa, T.; Sameshima, T.; Sato, Y.; Sumiyoshi, Y.; Ueno, T.; Shirasaki, Y.; Funatsu, T.; Shoji, Shuichi.

    12th International Conference on Miniaturized Systems for Chemistry and Life Sciences - The Proceedings of MicroTAS 2008 Conference. Chemical and Biological Microsystems Society, 2008. p. 296-298.

    Research output: Chapter in Book/Report/Conference proceedingConference contribution

    Arakawa, T, Sameshima, T, Sato, Y, Sumiyoshi, Y, Ueno, T, Shirasaki, Y, Funatsu, T & Shoji, S 2008, Dynamic imaging of single biomolecular interaction using flow control and TIRFM. in 12th International Conference on Miniaturized Systems for Chemistry and Life Sciences - The Proceedings of MicroTAS 2008 Conference. Chemical and Biological Microsystems Society, pp. 296-298, 12th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2008, San Diego, CA, 08/10/12.
    Arakawa T, Sameshima T, Sato Y, Sumiyoshi Y, Ueno T, Shirasaki Y et al. Dynamic imaging of single biomolecular interaction using flow control and TIRFM. In 12th International Conference on Miniaturized Systems for Chemistry and Life Sciences - The Proceedings of MicroTAS 2008 Conference. Chemical and Biological Microsystems Society. 2008. p. 296-298
    Arakawa, T. ; Sameshima, T. ; Sato, Y. ; Sumiyoshi, Y. ; Ueno, T. ; Shirasaki, Y. ; Funatsu, T. ; Shoji, Shuichi. / Dynamic imaging of single biomolecular interaction using flow control and TIRFM. 12th International Conference on Miniaturized Systems for Chemistry and Life Sciences - The Proceedings of MicroTAS 2008 Conference. Chemical and Biological Microsystems Society, 2008. pp. 296-298
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    AU - Sameshima, T.

    AU - Sato, Y.

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    AU - Ueno, T.

    AU - Shirasaki, Y.

    AU - Funatsu, T.

    AU - Shoji, Shuichi

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