Nature Biotechnology
23, 361 - 365 (2005)
Published online: 20 February 2005; | doi:10.1038/nbt1072
Microfabricated arrays of femtoliter chambers allow single molecule enzymologyYannick Rondelez1, Guillaume Tresset1, Kazuhito V Tabata2, 3, Hideyuki Arata2, Hiroyuki Fujita2, Shoji Takeuchi2
& Hiroyuki Noji21
Laboratory for Integrated Micro Mechatronic Systems (LIMMS-CNRS/IIS), University of Tokyo, 4-6-1 Komaba, Meguroku, 153-8505 Tokyo, Japan. 2
Institute of Industrial Science (IIS), University of Tokyo, 4-6-1 Komaba, Meguroku, 153-8505 Tokyo, Japan. 3
Research on Advanced Medical Technology, Japan Association for the Advancement of Medical Equipment (JAAME), 3-42-6, Hongo, Bunkyou-ku,
Tokyo, Japan.
Correspondence should be addressed to Hiroyuki Noji hnoji@iis.u-tokyo.ac.jpPrecise understanding of biological functions requires tools comparable in size to the basic components of life1,
2,
3,
4.
Single molecule studies have revealed molecular behaviors usually hidden in the ensemble- and time-averaging of bulk experiments5,
6. Although most such approaches rely on sophisticated optical strategies to limit the detection volume7,
8, another attractive approach is to perform the assay inside very small containers9,
10,
11,
12,
13,
14,
15,
16. We have developed a silicone device presenting a large array of micrometer-sized cavities. We used it to tightly enclose volumes of solution, as low as femtoliters, over long periods of time. The microchip insures that the chambers are uniform and precisely positioned. We demonstrated the feasibility of our approach by measuring the activity of single molecules of  -galactosidase and horseradish peroxidase. The approach should be of interest for many ultrasensitive bioassays at the single-molecule level.
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or 100 
-d-galactopyranoside (FDG), fluorescein, Bodipy492/515disulfonate, sulforhodamineG, SYBRGreen and fluorescent beads (Molecular Probes)
