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

Research output: Contribution to conferencePaper

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
Pages296-298
Number of pages3
Publication statusPublished - 2008 Jan 1
Event12th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2008 - San Diego, CA, United States
Duration: 2008 Oct 122008 Oct 16

Conference

Conference12th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2008
CountryUnited States
CitySan Diego, CA
Period08/10/1208/10/16

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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., Funatsu, T., & Shoji, S. (2008). Dynamic imaging of single biomolecular interaction using flow control and TIRFM. 296-298. Paper presented at 12th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2008, San Diego, CA, United States.