Monitoring enzymatic reactions with in situ sensors

Ian T. Young; V. Iordanov; Arthur Kroon; Heidi R. C. Dietrich; R. Moerman; L. Richard van den Doel; G.W. K. van Dedem; Andre Bossche; Bonnie L. Gray; Lina Sarro; Piet W. Verbeek; Lucas J. van Vliet

doi:10.1117/12.478088

18 July 2003 Monitoring enzymatic reactions with in situ sensors

Ian T. Young, V. Iordanov, Arthur Kroon, Heidi R. C. Dietrich, R. Moerman, L. Richard van den Doel, G.W. K. van Dedem, Andre Bossche, Bonnie L. Gray, Lina Sarro, Piet W. Verbeek, Lucas J. van Vliet

Author Affiliations +

Proceedings Volume 4966, Microarrays and Combinatorial Technologies for Biomedical Applications: Design, Fabrication, and Analysis; (2003) https://doi.org/10.1117/12.478088
Event: Biomedical Optics, 2003, San Jose, CA, United States

Abstract

In previous publications and presentations we have described our construction of a laboratory-on-a-chip based on nanoliter capacity wells etched in silicon. We have described methods for dispensing reagents as well as samples, for preventing evaporation, for embedding electronics in each well to measure fluid volume per well in real-time, and for monitoring the production or consumption of NADH in enzyme-catalyzed reactions such as those found in the glycolytic pathway of yeast. In this paper we describe the use of light sensors (photodiodes) in each well to measure both fluorescence (such as that evidenced in NADH) as well as bioluminescence (such as evidenced in ATP assays). We show that our detection limit for NADH fluorescence in 100 μM and for ATP/luciferase bioluminescence is 2.4 μM.

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Ian T. Young, V. Iordanov, Arthur Kroon, Heidi R. C. Dietrich, R. Moerman, L. Richard van den Doel, G.W. K. van Dedem, Andre Bossche, Bonnie L. Gray, Lina Sarro, Piet W. Verbeek, and Lucas J. van Vliet "Monitoring enzymatic reactions with in situ sensors", Proc. SPIE 4966, Microarrays and Combinatorial Technologies for Biomedical Applications: Design, Fabrication, and Analysis, (18 July 2003); https://doi.org/10.1117/12.478088

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