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µ-RA: Micro reactor arrays for high-throughput and ultra-sensitive bioassays
Understanding how bio-molecules organize and function in a cell will require tools comparable in size to the basic components of life. It means the tools must be in the ‘right’ size to handle these bio-molecules at biologically relevant scale. Nanotechnologies in combination with microscopic techniques can allow the analysis of nanometer-sized bio-molecules inside very small containers (reactors).
We, at Ichiki laboratory (University of Tokyo), have developed a microfabricated chip using photolithography which is comprising of uniformly distributed femtoliter scale reactors in 10,000 microchabmber/mm2. The diameter and volume of single micro chambers are ~5µm and ~120 femtoliters, respectively (see Fig. below).
We have evaluated the enzymatic activity of mutant aldehyde reductase (AKR) inside the micro reactors in a sub-picoliter scale which corresponds to 10,000 molecules per reactor. We also evaluated and demonstrated the feasibility of single-molecule level enzyme assay for beta-galactosidase inside the micro reactor.
We expect this tool will be useful in a number of biomedical applications where accurate and highly sensitive target analysis is required.
References:
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Manish Biyani, Terutsune Osawa, Naoto Nemoto and Takanori Ichiki (2008) One-to-one gene-encoded functional protein microarray. In: Proc of 12th International Conference on Miniaturized Systems for
Chemistry and Life Sciences. MicroTAS, San Diego, USA, vol.2, p.p.1781-1783.
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Manish Biyani, Naoto Nemoto, Yosuke Hosoi and Takanori Ichiki (2006) RNA-to-Protein Chip:
an integrated microarray platform to bridge genomics and proteomics. In: Proc. of 10th
International Conference on Miniaturized Systems for Chemistry and Life Sciences. MicroTAS, Tokyo, Japan, vol.1, p.p.209-211
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Yosuke Hosoi, Manish Biyani, Naoto Nemoto, Takanori Akagi and Takanori Ichiki (2005)
High-throughput screening of mutant biomolecules using mRNA display and microreactor array chips. In:
Proc. of International Conference on Solid State Device and Materials, SSDM, Kobe, Japan, pp.452-453.
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Yosuke Hosoi, Manish Biyani, Naoto Nemoto and Takanori Ichiki (2005) High-Throughput Screening of mutant AKR enzymes using mRNA display and novel microreactor array chips. In: Proc. of 9th
International Conference on Miniaturized Systems for Chemistry and Life Sciences,
MicroTAS, Boston, USA, vol.1, p.p. 323-325.
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