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Microfabricated arrays of femtoliter chambers allow single molecule enzymology

Microfabricated arrays of femtoliter chambers allow single molecule enzymology Precise understanding of biological functions requires tools comparable in size to the basic components of life 1,2,3,4 . Single molecule studies have revealed molecular behaviors usually hidden in the ensemble- and time-averaging of bulk experiments 5,6 . Although most such approaches rely on sophisticated optical strategies to limit the detection volume 7,8 , another attractive approach is to perform the assay inside very small containers 9,10,11,12,13,14,15,16 . 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 β-galactosidase and horseradish peroxidase. The approach should be of interest for many ultrasensitive bioassays at the single-molecule level. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Nature Biotechnology Springer Journals

Microfabricated arrays of femtoliter chambers allow single molecule enzymology

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References (36)

Publisher
Springer Journals
Copyright
Copyright © 2005 by Nature Publishing Group
Subject
Life Sciences; Life Sciences, general; Biotechnology; Biomedicine, general; Agriculture; Biomedical Engineering/Biotechnology; Bioinformatics
ISSN
1087-0156
eISSN
1546-1696
DOI
10.1038/nbt1072
Publisher site
See Article on Publisher Site

Abstract

Precise understanding of biological functions requires tools comparable in size to the basic components of life 1,2,3,4 . Single molecule studies have revealed molecular behaviors usually hidden in the ensemble- and time-averaging of bulk experiments 5,6 . Although most such approaches rely on sophisticated optical strategies to limit the detection volume 7,8 , another attractive approach is to perform the assay inside very small containers 9,10,11,12,13,14,15,16 . 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 β-galactosidase and horseradish peroxidase. The approach should be of interest for many ultrasensitive bioassays at the single-molecule level.

Journal

Nature Biotechnology – Springer Journals

Published: Feb 20, 2005

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