The VIIBRE Automated Biosystems Core (ABC) provides the University and its collaborating institutions with a powerful technical foundation for the study of dynamic cellular metabolism, signaling, and control. The instruments were acquired or built with more than $4.7 million of specific funding received from the Defense Threat Reduction Agency (DTRA), the National Institute on Drug Abuse (NIDA), the NIH S10 Shared Instrumentation Program, and Vanderbilt University. A major focus of the ABC is to explore dynamic host responses to drugs, chemical agents, toxins, and microbial pathogens by offering acquisition of multi-parameter quantitative data sets of cell populations, individual cells, and the cellular secretome with great statistical validation power, automated analysis routines that discern small changes, and linked events. The ABC operates two major instrument groups, the first for automated high-content imaging and the second for cellular omni-omic characterization.
The first ABC group includes a microfluidics-enabled high-content screening (HCS) system built around a Perkin Elmer Opera QEHS Automated Confocal Microscopy System. The PerkinElmer Opera QEHS is a state-of-the-art, high-throughput, high-content confocal screening microscope. It provides ultra-fast, high-resolution imaging of 96, 384, and 1536 wellplates, glass slides, or VIIBRE's custom microfluidic wellplates and zebrafish holders. The instrument is capable of acquiring about 80 to 400+ images (1.3 MP, 12-bit confocal) per minute, depending on imaging setup. It utilizes up to 4 CCD cameras (3 Peltier-cooled), 4 lasers (405, 488, 561, 640 nm) plus a tunable UV/Vis source and TLED-brightfield illumination. The confocal spinning-disk technique with 40 ms minimum exposure times allows for minimal bleaching of samples, while taking high-quality images at very high frame rates, which makes it ideal for live cell imaging. A multitude of optical filters and the option to separate or combine exposures meet a wide range of fluorescent labeling needs.
The second ABC instrument group is a custom Omni-Omics system built around a Waters Synapt G2 Ion Mobility-Mass Spectrometer (IM-MS) and an automated Nikon Eclipse microscope and other real-time instruments and microfabricated bioreactors for sensing and controlling small populations of cells in microfluidic devices. The IM-MS has nanoelectrospray (nESI), matrix-assisted laser desorption ionization (MALDI), and ultraperformance liquid chromatography (UPLC) and gas chromatography (GC) sample preprocessing with automated sample handling. For operating in nESI mode, an automated chromatographic desalting system uses a four-channel Exigent HPLC pump and allows us to conduct a proteomic / metabolomic screen every ten minutes – a major advance enabled by the replacement of slow, liquid-phase high-performance liquid chromatography with the millisecond separations provided by a gas phase-ion mobility system. This IM-MS system is also supported by a fully automated, Nikon Eclipse inverted fluorescence microscope with a long-travel sub-micron resolution stage interfaced to an attached fluid-handling robot; real-time electrochemical sensing of cellular metabolites, including glucose and oxygen consumption, lactate production, and acidification; custom 96-wellplates with internal microfabricated cell traps and microfluidic perfusion control; and a computer-controlled microliter formulator to create microliter volumes of custom perfusion solutions in a minute or two.
For questions regarding grant information, Screening4Hire, training, and assay setup or to schedule the instrument, please contact ABCmanager@vanderbilt.edu.