Fluorescent and Luminescent Kinetic Imaging Plate Reader

Lindsley, Craig

Abstract

Vanderbilt University is requesting funds for the purchase of a Wave Front Biosciences Panoptic kinetic imaging plate reader to replace the obsolete Hamamatsu FDSS 6000. Kinetic imaging plate readers are designed to simultaneously image every well of entire multi-well assay plates at high speeds in order to monitor changes in cellular and cell-free systems over time. Assays that most commonly use kinetic imaging plate readers are fluorescence/luminescence-based assays of intracellular ions (e.g. Ca2+), cyclic nucleotides, membrane potential, as well as cell-free enzyme-mediated substrate-product conversion. Vanderbilt's FDSS was purchased in 2004 using funds awarded as a S10 shared instrumentation grant. The FDSS was installed in the Vanderbilt High-throughput Screening Facility where it has served as the cornerstone for Vanderbilt's small-molecule probe and early stage drug discovery efforts. Over the course of the last ten years, the FDSS has been by far the most used plate reader in the HTS Facility where its use has resulted in over 40 publications, over 30 patents, and over 5 licensing agreement with pharmaceutical companies and has supported over 50 individual research projects ranging from neuroscience, metabolic diseases, anti-microbials, cancer, and insect-vectors of disease. Although the FDSS has been an outstanding instrument, it is past its life expectancy, is no longer produced by Hamamatsu, and, critically, after this year will no longer be supported by Hamamatsu. Thus, it is crucial that this instrument is replaced as soon as possible to avoid crippling numerous NIH-funded as well as burgeoning research programs aimed at advancing basic and early translational research. After careful consideration, we are requesting funds for the purchase of the Panoptic instead of the present version of the FDSS, FDSS 7000, or the competing FLIPR Tetra based on the Panoptic's superior combination of features and their ability to address the present and future needs of the Vanderbilt research community who depend on the kinetic imaging plate reader for their research. In particular: 1) Panoptic's ability to seamlessly measure ultra-low light signals from luminescent reporters such as Promega's glosensor and scintillation proximity assays to high- speed imaging or bright fluorescent signals 2) simple, rapid, and economical adaptation of a wide-variety of assays systems presently in use or under development including multi-channel/multiplexed assay, BRET, FRET, and fluorescence polarization through the use 10 position and 5 position high-speed excitation and emission filter changers 3) vastly more flexible liquid handling, modular peripherals, and advanced automation interface to enable facile adaptation of the kinetic imaging plate reader to a growing list of assay systems. The timely replacement of the obsolete FDSS 6000 with the considerably more capable Panoptic will ensure uninterrupted service to presently funded projects while providing a means to advance the use of kinetic imaging plate readers to a wider variety of biological systems and assay formats.

Public Health Relevance

In order to study rapidly occurring physiological process that are of critical importance to human health, such as neurotransmission and cardiac muscle contraction, researchers require high-speed imaging systems. Traditionally microscopes have been a mainstay of this research, however microscopes are very slow compared to kinetic imaging plate readers which are capable increasing the number of simultaneous experiments hundreds of fold. The increase in experimental throughput afforded by kinetic imaging plate readers will enable researchers to dramatically advance our understanding of rapid physiological processes which, in turn, will lead to more effective development of new therapies for disease.