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Time-resolved fluorescence measurements using microlens array and area imaging devices.

Susanne Merk1, Achim Lietz, Margareta Kroner

  • 1Boehringer Ingelheim Pharma GmbH & Co. KG, Department of Integrated Lead Discovery, Birkendorfer Strasse 65, D-88397 Biberach an der Riss, Germany.

Combinatorial Chemistry & High Throughput Screening
|February 18, 2004
PubMed
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The Plate::Vision reader offers superior sensitivity for time-resolved fluorescence (TRF) assays compared to the LEADseeker, achieving lower detection limits for europium and better performance in TRF resonance energy transfer (TR-FRET) experiments, enhancing high-throughput screening capabilities.

Area of Science:

  • Biotechnology
  • Assay Development
  • High-Throughput Screening

Background:

  • Time-resolved fluorescence (TRF) assay formats are crucial for high-throughput screening (HTS).
  • Novel imaging systems aim to improve sensitivity and throughput in TRF applications.

Purpose of the Study:

  • To characterize and compare the performance of the Plate::Vision 96-microlens array reader and the LEADseeker Generation IV imaging system.
  • To evaluate their suitability for TRF and TRF resonance energy transfer (TR-FRET) assay modes.

Main Methods:

  • Europium measurements were conducted to determine the limit of detection (LOD) for both readers.
  • TR-FRET experiments involved titrating europium-biotin (Eu-biotin) with streptavidin-allophycocyanin (SA-APC) to assess Z' values.
  • A drug screening simulation using TR-FRET measured energy transfer efficiency.

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Main Results:

  • Plate::Vision demonstrated a lower LOD for europium (approx. 3 pM) compared to LEADseeker (approx. 10 pM).
  • Plate::Vision achieved higher Z' values (>0.5) at lower concentrations in TR-FRET assays than LEADseeker.
  • Both systems offer increased throughput due to reduced measurement times compared to traditional readers.

Conclusions:

  • Plate::Vision exhibits superior sensitivity and performance for TRF and TR-FRET applications compared to LEADseeker.
  • Both novel readers provide good quality results and enhance throughput for TRF-based screening.
  • These systems represent advancements in TRF assay technologies for drug discovery.