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Journal of Biomolecular Screening
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Homogeneous Cell- and Bead-Based Assays for High Throughput Screening Using Fluorometric Microvolume Assay Technology

Sheri Miraglia

PE Biosystems, Foster City, CA

Elana E. Swartzman

PE Biosystems, Foster City, CA

Julia Mellentin-Michelotti

PE Biosystems, Foster City, CA

Lolita Evangelista

PE Biosystems, Foster City, CA

Christopher Smith

PE Biosystems, Foster City, CA

Iwan Gunawan

PE Biosystems, Foster City, CA

Kenton Lohman

Biometric Imaging, Mountain View, CA

Edward M. Goldberg

Molecular Epidemiology Laboratory, Institute for Environmental Safety and Occupational Health Risk Assessment, Brooks AFB, San Antonio, TX

Bala Manian

Molecular Epidemiology Laboratory, Institute for Environmental Safety and Occupational Health Risk Assessment, Brooks AFB, San Antonio, TX

Pau-Miau Yuan

PE Biosystems, Foster City, CA

High throughput drug screening has become a critical component of the drug discovery process. The screening of libraries containing hundreds of thousands of compounds has resulted in a requirement for assays and instrumentation that are amenable to nonradioactive formats and that can be miniaturized. Homogeneous assays that minimize upstream automation of the individual assays are also preferable. Fluorometric microvolume assay technology (FMAT) is a fluorescence-based platform for the development of nonradioactive cell- and bead-based assays for HTS. This technology is plate format-independent, and while it was designed specifically for homogeneous ligand binding and immunological assays, it is amenable to any assay utilizing a fluorescent cell or bead. The instrument fits on a standard laboratory bench and consists of a laser scanner that generates a 1 mm2 digitized image of a 100-µm deep section of the bottom of a microwell plate. The instrument is directly compatible with a Zymark TwisterTM (Zymark Corp., Hopkinton, MA) for robotic loading of the scanner and unattended operation in HTS mode. Fluorescent cells or beads at the bottom of the well are detected as localized areas of concentrated fluorescence using data processing. Unbound flurophore comprising the background signal is ignored, allowing for the development of a wide variety of homogeneous assays. The use of FMAT for peptide ligand binding assays, immunofluorescence, apoptosis and cytotoxicity, and bead-based immunocapture assays is described here, along with a general overview of the instrument and software.

Journal of Biomolecular Screening, Vol. 4, No. 4, 193-204 (1999)
DOI: 10.1177/108705719900400407
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