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; S. Shoji

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, S. Shoji

School of Fundamental Science and Engineering

Research output: Contribution to conference › Paper › peer-review

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 language	English
Pages	296-298
Number of pages	3
Publication status	Published - 2008 Jan 1
Event	12th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2008 - San Diego, CA, United States Duration: 2008 Oct 12 → 2008 Oct 16

Conference

Conference	12th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2008
Country/Territory	United States
City	San Diego, CA
Period	08/10/12 → 08/10/16

Keywords

Microfluidic device
Single biomolecular imaging
TIRFM

ASJC Scopus subject areas

Chemical Engineering (miscellaneous)
Bioengineering

Cite this

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title = "Dynamic imaging of single biomolecular interaction using flow control and TIRFM",

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.",

keywords = "Microfluidic device, Single biomolecular imaging, TIRFM",

author = "T. Arakawa and T. Sameshima and Y. Sato and Y. Sumiyoshi and T. Ueno and Y. Shirasaki and T. Funatsu and S. Shoji",

year = "2008",

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language = "English",

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T1 - Dynamic imaging of single biomolecular interaction using flow control and TIRFM

AU - Arakawa, T.

AU - Sameshima, T.

AU - Sato, Y.

AU - Sumiyoshi, Y.

AU - Ueno, T.

AU - Shirasaki, Y.

AU - Funatsu, T.

AU - Shoji, S.

PY - 2008/1/1

Y1 - 2008/1/1

N2 - 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.

AB - 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.

KW - Microfluidic device

KW - Single biomolecular imaging

KW - TIRFM

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M3 - Paper

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T2 - 12th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2008

Y2 - 12 October 2008 through 16 October 2008

ER -

Dynamic imaging of single biomolecular interaction using flow control and TIRFM

Abstract

Conference

Keywords

ASJC Scopus subject areas

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