Rapid multi-reagents exchange TIRFM microfluidic system for single biomolecular imaging

Takahiro Arakawa, Tomoya Sameshima, Yukiko Sato, Taro Ueno, Yoshitaka Shirasaki, Takashi Funatsu, Shuichi Shoji

    Research output: Contribution to journalArticle

    9 Citations (Scopus)

    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. This TIRFM microfluidic system was constructed by a smooth glass in microchannel surface, that is essential for TIR imaging, and the polydimethylesiloxane (PDMS) microvalves that is operatable with a rapid response of around 100 ms. The smooth glass surface was etched with an optimized hydrofluoric acid and nitric acid (HF-HNO3) solution and had the average surface roughness of 3.2 nm. The microvalves were based on a pneumatic PDMS membrane that is beneficial for a rapid (<100 ms) response. This TIRFM microfluidic system contains three inlet channels and one outlet channel and the flow in every inlet channel were precisely controlled by the PDMS microvalves. These microvalves were operated by air pressure pulses of 100 ms to hold, open or closed for switching. This TIRFM microfluidic system enabled a reagent exchange within 100 ms with TIRFM observation. 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. The micro-constructed TIRFM microfluidic system can realize dynamic analysis of the single molecule level protein-protein binding and dissociation under a controlled multi-reagent exchange.

    Original languageEnglish
    Pages (from-to)218-225
    Number of pages8
    JournalSensors and Actuators, B: Chemical
    Volume128
    Issue number1
    DOIs
    Publication statusPublished - 2007 Dec 12

    Fingerprint

    Fluorescence microscopy
    Microfluidics
    reagents
    microscopy
    Imaging techniques
    fluorescence
    Microchannels
    Glass
    microchannels
    proteins
    Fluorescence
    Chaperonins
    glass
    Hydrofluoric Acid
    Proteins
    Molecular imaging
    Nitric Acid
    Hydrofluoric acid
    Molecules
    Streptavidin

    Keywords

    • GroEL
    • GroES
    • Microvalve
    • Multi-reagent exchange
    • PDMS
    • Single biomolecular imaging
    • TIRFM microfluidic system

    ASJC Scopus subject areas

    • Analytical Chemistry
    • Electrochemistry
    • Electrical and Electronic Engineering

    Cite this

    Rapid multi-reagents exchange TIRFM microfluidic system for single biomolecular imaging. / Arakawa, Takahiro; Sameshima, Tomoya; Sato, Yukiko; Ueno, Taro; Shirasaki, Yoshitaka; Funatsu, Takashi; Shoji, Shuichi.

    In: Sensors and Actuators, B: Chemical, Vol. 128, No. 1, 12.12.2007, p. 218-225.

    Research output: Contribution to journalArticle

    Arakawa, Takahiro ; Sameshima, Tomoya ; Sato, Yukiko ; Ueno, Taro ; Shirasaki, Yoshitaka ; Funatsu, Takashi ; Shoji, Shuichi. / Rapid multi-reagents exchange TIRFM microfluidic system for single biomolecular imaging. In: Sensors and Actuators, B: Chemical. 2007 ; Vol. 128, No. 1. pp. 218-225.
    @article{123cac505a3341df95101bd8607f90e6,
    title = "Rapid multi-reagents exchange TIRFM microfluidic system for single biomolecular imaging",
    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. This TIRFM microfluidic system was constructed by a smooth glass in microchannel surface, that is essential for TIR imaging, and the polydimethylesiloxane (PDMS) microvalves that is operatable with a rapid response of around 100 ms. The smooth glass surface was etched with an optimized hydrofluoric acid and nitric acid (HF-HNO3) solution and had the average surface roughness of 3.2 nm. The microvalves were based on a pneumatic PDMS membrane that is beneficial for a rapid (<100 ms) response. This TIRFM microfluidic system contains three inlet channels and one outlet channel and the flow in every inlet channel were precisely controlled by the PDMS microvalves. These microvalves were operated by air pressure pulses of 100 ms to hold, open or closed for switching. This TIRFM microfluidic system enabled a reagent exchange within 100 ms with TIRFM observation. 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. The micro-constructed TIRFM microfluidic system can realize dynamic analysis of the single molecule level protein-protein binding and dissociation under a controlled multi-reagent exchange.",
    keywords = "GroEL, GroES, Microvalve, Multi-reagent exchange, PDMS, Single biomolecular imaging, TIRFM microfluidic system",
    author = "Takahiro Arakawa and Tomoya Sameshima and Yukiko Sato and Taro Ueno and Yoshitaka Shirasaki and Takashi Funatsu and Shuichi Shoji",
    year = "2007",
    month = "12",
    day = "12",
    doi = "10.1016/j.snb.2007.06.014",
    language = "English",
    volume = "128",
    pages = "218--225",
    journal = "Sensors and Actuators, B: Chemical",
    issn = "0925-4005",
    publisher = "Elsevier",
    number = "1",

    }

    TY - JOUR

    T1 - Rapid multi-reagents exchange TIRFM microfluidic system for single biomolecular imaging

    AU - Arakawa, Takahiro

    AU - Sameshima, Tomoya

    AU - Sato, Yukiko

    AU - Ueno, Taro

    AU - Shirasaki, Yoshitaka

    AU - Funatsu, Takashi

    AU - Shoji, Shuichi

    PY - 2007/12/12

    Y1 - 2007/12/12

    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. This TIRFM microfluidic system was constructed by a smooth glass in microchannel surface, that is essential for TIR imaging, and the polydimethylesiloxane (PDMS) microvalves that is operatable with a rapid response of around 100 ms. The smooth glass surface was etched with an optimized hydrofluoric acid and nitric acid (HF-HNO3) solution and had the average surface roughness of 3.2 nm. The microvalves were based on a pneumatic PDMS membrane that is beneficial for a rapid (<100 ms) response. This TIRFM microfluidic system contains three inlet channels and one outlet channel and the flow in every inlet channel were precisely controlled by the PDMS microvalves. These microvalves were operated by air pressure pulses of 100 ms to hold, open or closed for switching. This TIRFM microfluidic system enabled a reagent exchange within 100 ms with TIRFM observation. 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. The micro-constructed TIRFM microfluidic system can realize dynamic analysis of the single molecule level protein-protein binding and dissociation under a controlled multi-reagent exchange.

    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. This TIRFM microfluidic system was constructed by a smooth glass in microchannel surface, that is essential for TIR imaging, and the polydimethylesiloxane (PDMS) microvalves that is operatable with a rapid response of around 100 ms. The smooth glass surface was etched with an optimized hydrofluoric acid and nitric acid (HF-HNO3) solution and had the average surface roughness of 3.2 nm. The microvalves were based on a pneumatic PDMS membrane that is beneficial for a rapid (<100 ms) response. This TIRFM microfluidic system contains three inlet channels and one outlet channel and the flow in every inlet channel were precisely controlled by the PDMS microvalves. These microvalves were operated by air pressure pulses of 100 ms to hold, open or closed for switching. This TIRFM microfluidic system enabled a reagent exchange within 100 ms with TIRFM observation. 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. The micro-constructed TIRFM microfluidic system can realize dynamic analysis of the single molecule level protein-protein binding and dissociation under a controlled multi-reagent exchange.

    KW - GroEL

    KW - GroES

    KW - Microvalve

    KW - Multi-reagent exchange

    KW - PDMS

    KW - Single biomolecular imaging

    KW - TIRFM microfluidic system

    UR - http://www.scopus.com/inward/record.url?scp=35748964432&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=35748964432&partnerID=8YFLogxK

    U2 - 10.1016/j.snb.2007.06.014

    DO - 10.1016/j.snb.2007.06.014

    M3 - Article

    VL - 128

    SP - 218

    EP - 225

    JO - Sensors and Actuators, B: Chemical

    JF - Sensors and Actuators, B: Chemical

    SN - 0925-4005

    IS - 1

    ER -