Microinjection of sigma-D-glucose standards and Amplex Red reagent on micro-arrays

R. Moerman; L. Richard van den Doel; S. Picioreanu; J. Frank; Johannes P. A. Marijnissen; G.W. K. van Dedem; Kari H. Hjelt; Michael J. Vellekoop; Pasqualina M. Sarro; Ian T. Young

doi:10.1117/12.350052

3 June 1999 Microinjection of sigma-D-glucose standards and Amplex Red reagent on micro-arrays

R. Moerman, L. Richard van den Doel, S. Picioreanu, J. Frank, Johannes P. A. Marijnissen, G.W. K. van Dedem, Kari H. Hjelt, Michael J. Vellekoop, Pasqualina M. Sarro, Ian T. Young

Author Affiliations +

Proceedings Volume 3606, Micro- and Nanofabricated Structures and Devices for Biomedical Environmental Applications II; (1999) https://doi.org/10.1117/12.350052
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

Abstract

Intelligent Molecular Diagnostic Systems (IMDS)- The objective of this multidisciplinary research program is to design and develop an analytical system that is able to measure and interpret concentrations of various analytes which are dispensed on a micro-array. The analytes are detected by means of fluorescence or (chemi)luminescence measurement. Furthermore, the collected data are combined and interpreted using modern reasoning techniques. Micro-injection- Dispensing picoliters (pl) of reagents (enzymes, antibodies, etc.) and liquid samples on a micro-array requires special techniques. At the moment we are working on a technique which will allow for accurately dispensing liquid volumes less than 100 pl on a micro-array. Detection of (beta) -D-glucose- (beta) -D-glucose standards are dispensed on a micro-array, after which a solution of Amplex Red reagent, horse radish peroxidase (HARP), and glucose oxidase in a mixture of ethylene glycol and water is added. Ethylene glycol is added to prevent evaporation. The (beta) -D-glucose reacts with glucose oxidase to D-gluconolactone and H₂O₂. The H₂O₂ reacts with 10-acetyl-3,7-dihydroxyphenoxazine (Amplex Red) with a 1:1 stoichiometry to produce highly fluorescent resorufin. The formation of resorufin with time is followed with a Zeiss Axioskop microscope equipped with a KAF Photometrics CCD camera, in order to determine the sensitivity, concentrations, and volumes associated with the dispensed fluids.

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R. Moerman, L. Richard van den Doel, S. Picioreanu, J. Frank, Johannes P. A. Marijnissen, G.W. K. van Dedem, Kari H. Hjelt, Michael J. Vellekoop, Pasqualina M. Sarro, and Ian T. Young "Microinjection of sigma-D-glucose standards and Amplex Red reagent on micro-arrays", Proc. SPIE 3606, Micro- and Nanofabricated Structures and Devices for Biomedical Environmental Applications II, (3 June 1999); https://doi.org/10.1117/12.350052

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