Time-resolved UV-excited microarray reader for fluorescence energy transfer (FRET) measurements

Adelina Orellana; Ari Petteri Hokkanen; Tomi Pastinen; Kristina Takkinen; Hans Soderlund

doi:10.1117/12.426728

10 May 2001 Time-resolved UV-excited microarray reader for fluorescence energy transfer (FRET) measurements

Adelina Orellana, Ari Petteri Hokkanen, Tomi Pastinen, Kristina Takkinen, Hans Soderlund

Proceedings Volume 4252, Advances in Fluorescence Sensing Technology V; (2001) https://doi.org/10.1117/12.426728
Event: BiOS 2001 The International Symposium on Biomedical Optics, 2001, San Jose, CA, United States

Abstract

Analytical systems based on immunochemistry are largely used in medical diagnostics and in biotechnology. There is a significant pressure to develop the present assay formats to become easier to use, faster, and less reagent consuming. Further developments towards high density array--like multianalyte measurement systems would be valuable. To this aim we have studied the applicability of fluorescence resonance energy transfer and time-resolved fluorescence resonance energy transfer in immunoassays on microspots and in microwells. We have used engineered recombinant antibodies detecting the pentameric protein CRP as a model analyte system, and tested different assay formats. We describe also the construction of a time-resolved scanning epifluorometer with which we could measure the FRET interaction between the slow fluorescence decay from europium chelates and its energy transfer to the rapidly decaying fluorophore Cy5.

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Adelina Orellana, Ari Petteri Hokkanen, Tomi Pastinen, Kristina Takkinen, and Hans Soderlund "Time-resolved UV-excited microarray reader for fluorescence energy transfer (FRET) measurements", Proc. SPIE 4252, Advances in Fluorescence Sensing Technology V, (10 May 2001); https://doi.org/10.1117/12.426728

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KEYWORDS

Fluorescence resonance energy transfer

Luminescence

Glasses

Europium

Proteins

Photomultipliers

Energy transfer

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