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Proceedings Paper

Fluorescence detection in (sub-)nanoliter microarrays
Author(s): L. Richard van den Doel; Michael J. Vellekoop; Pasqualina M. Sarro; S. Picioreanu; R. Moerman; J. Frank; G.W. K. van Dedem; Kari H. Hjelt; Lucas J. van Vliet; Ian T. Young
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Paper Abstract

The goal of our TU Delft interfaculty research program is to develop intelligent molecular diagnostic systems (IMDS) that can analyze liquid samples that contain a variety of biochemical compounds such as those associated with fermentation processes. One specific project within the IMDS program focuses on photon sensors. In order to analyze the liquid samples we use dedicated microarrays. At this stage, these are basically miniaturized micro titre plates. Typical dimensions of a vial are 200 X 200 X 20 micrometer3. These dimensions may be varied and the shape of the vials can be modified with a result that the volume of the vials varies from 0.5 to 1.6 nl. For all experiments, we have used vials with the shape of a truncated pyramid. These vials are fabricated in silicon by a wet etching process. For testing purposes the vials are filled with rhodamine solutions of various concentrations. To avoid evaporation glycerol-water (1:1, v/v) with a viscosity of 8.3 times the viscosity of water is used as solvent. We aim at wide field-of-view imaging at the expense of absolute sensitivity: the field-of-view increases quadratically with decreasing magnification. Small magnification, however, implies low Numerical Aperture (NA). The ability of a microscope objective to collect photons is proportional to the square of the NA. To image the entire microarray we have used an epi-illumination fluorescence microscope equipped with a low magnification (2.5 X/0.075) objective and a scientific CCD camera to integrate the photons emitted from the fluorescing particles in the solutions in the vials. From these experiments we found that for this setup the detection limit is on the order of micromolar concentrations of fluorescing particles. This translates to 108 molecules per vial.

Paper Details

Date Published: 3 June 1999
PDF: 12 pages
Proc. SPIE 3606, Micro- and Nanofabricated Structures and Devices for Biomedical Environmental Applications II, (3 June 1999); doi: 10.1117/12.350059
Show Author Affiliations
L. Richard van den Doel, Delft Univ. of Technology (Netherlands)
Michael J. Vellekoop, Delft Univ. of Technology (Netherlands)
Pasqualina M. Sarro, Delft Univ. of Technology (Netherlands)
S. Picioreanu, Delft Univ. of Technology (Netherlands)
R. Moerman, Delft Univ. of Technology (Netherlands)
J. Frank, Delft Univ. of Technology (Netherlands)
G.W. K. van Dedem, Delft Univ. of Technology (Netherlands)
Kari H. Hjelt, Delft Univ. of Technology (Netherlands)
Lucas J. van Vliet, Delft Univ. of Technology (Netherlands)
Ian T. Young, Delft Univ. of Technology (Netherlands)

Published in SPIE Proceedings Vol. 3606:
Micro- and Nanofabricated Structures and Devices for Biomedical Environmental Applications II
Mauro Ferrari, Editor(s)

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