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Prceedings of 2006 In ternational Conf erence on Microtechnologies in Medicine and Biology Okinawa, Japan 9-1 2 May 2006

Development of Biosensor Chip for Clinical Diagnosis Using Surface Plasmon Resonance Imaging with Multi-Microchannels
T. Suzuki, Y. Teramura*, K. Inokuma, I. Kanno, H. Iwata** and H. Kotera Dept. of Microengineering, Kyoto University Yoshida-honmachi, Sakyo-ku, Kyoto, 606-8501, Japan Tel +81-75-753-3559, Fax +81-75-771-7286, E-mail takaakigme.kyoto-u.ac.jp * Dept. of Polymer Chemistry, Kyoto University 53 Kawara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan * * Institute for Frontier Medical Sciences, Kyoto University 53 Kawara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan Abstract A compact biosensor chip for clinical diagnosis is presented. The proposed biochip integrated three independent microchannels on one chip partially coated with Au thin film as a Surface Plasmon Resonance (SPR) excitation layer. For clinical diagnosis, the affinity binding of unlabeled biological molecules onto the Au surface can be quantitatively analyzed by SPR imaging with the multi-microchannels, i.e. one biosample and two reference flows to obtain an analytical curve.

icrochannels Keyword: Surface Plasmon Resonance Imaging, Clinical Diagnosis, Multi-m

1 INTRODUCTION

Miniaturized SPR sensors with multichannels have been developed as an alternative to laboratory SPR systems to realize mobile, compact, and cost-effective sensing devices [1-3]. However, most of the systems in which microchannels and substrates are not bonded decrease the detection sensitivity due to cross contamination and dehydration of immobilized materials. In this paper, a compact and high-sensitive biochip for clinical diagnosis is presented. The proposed biochip are composed of three independent microchannels on one chip, and quantitatively measures the affinity binding of unlabeled biological molecules using the Surface Plasmon Resonance (SPR) imaging. The protein adsorption by the change in the microchannel height and the flow velocity was observed for geometry optimization of microchannels in the chip in order to improve the detection sensitivity and the throughput of

screenings.

20pm on a glass substrate. A master of microchannels was fabricated on the glass substrate using photolithography procedure. Then, liquid PDMS prepolymer was poured on the master and degassed using vacuum-forming to mold the microchannels. The PDMS microchannels were peeled off the master. On the other hand, thin Cr and Au films as a SPR excitation layer were partially deposited on glass substrates (refractive index n=1.52) using a vacuum evaporator with a metal mask. Finally, the PDMS microchannels and the glass substrates were bonded using 2 plasma bonding technique [5] in order to increase the detection sensitivity of the SPR imaging. Irreversible bonding increases upper limit of fluid velocity and liquid switching speed without cross contamination and dehydration of immobilized materials. The dimensions of the fabricated multi-microchannels were 500pm wide, 20pm height, and 2mm intervals. The biosensor chips can be used by putting on the optical apparatus, thus allowing it to be used as a low-cost disposable device.
2.2 Measurement

2 EXPERIMENTAL 2.1 Fabrication Process

Sensing microchannels based on the SPR imaging is rapidly and easily made from poly(dimethylsiloxane) (PDMS) using soft micromachining techniques as shown in Fig. 1. The thick photoresist SU-8 was first spin coated with a thickness of

To demonstrate sensing characteristics of the proposed chip, the adsorption of bovine serum albumin (BSA) onto the gold surface was evaluated by putting the prepared chip on an optical apparatus as shown in Fig.2. Three samples with different concentrations of BSAlwater were simultaneously infused to the microchannels by syringe pumps connected to inlets of microchanmles by Teflon microtubes.

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3 RESULTS AND DISCUSSION

3.1 Evaluation of Quantitative Measuremenit

interactions, e.g. detection of the tuimor marker im clinical diagnosis using the anten-antibody reaction.
3.2 Microchannel Height and Flow Velocity Dependlences

Three samples with 0.1%, 0.5% and l.0% BSA/water were simultaneously iMnfsed to thi fabricated microchannels. SPR images as a fuintion of time were clearly bserved by a CCD camera as shown in Fig.3. From SPR sensorgam denrved by the images m Fig.4, it is seen that the quantity an the rate of the adsorption depend on the concenrtio6n of thi sample im accordance with ldinetics. Therefore, the proposed system is apphcable ftr chlaracteiing and quaifyin biomblecular

We evaluated the protein adsorption by the change im the microchannel heig adi the flow velocity for geometry optimization of multi-microchaneIs in SPR iaing sensors in orde to improve the detection sensitivity nd the throhput of screenings. hi consequence, the adsorption rate is proportional to one-third power of the flow velocity, and is inversely proportional to one-third power of the heigt.

(2-a) t7hini Cr 1aerV with Im thicknev, iJ paftially opofttd on a gIas sUbWtrac using vacuum evaportinM

2-b) Thin Au laayr with 49nni thickness is partially deposited on a4 glass substratoe Osing vacuum evaporation

M prepolymnr is poured onto PMS (I c LNiOPid te MOld deaetated and cured,

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ACKNOWLEDGEMENTS

This study is a part of Kyoto City Collaboration of Regional Entities for the Advancement of Tecniological Excellence of JST on the basis of research results suported in part by Mizuho Foundation for the Promnotion of Sciences. and Center of Excellence for Research and Education on Complex Functional Mechanical Systems (COE program) of MEXT, Japan.
REFERENCES (b) 8min (a) Omin Fig. 3 SPR images as a fnction of time.
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Fig.4 SPR sen:sorgram of BSA/water at flow velocity 3.6mmlsee.

Time [mi]

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