Laboratory:
The high-throughput screening (HTS) facility, includes >2000 square feet of space designed to
support industrial-scale laboratory automation. In addition, there is a ~ 400 sq ft cell culture
facility adjacent to the main HTS lab. The HTS facility is fully-equipped with state-of-the art
readers, liquid handling, laboratory automation, and laboratory information management
systems (LIMS) needed to perform the complex cell-based and cell-free assays to support
Vanderbilt investigators in basic and translational research.
Major Equipment: (if too long, you may list with no description)
Vanderbilt has invested heavily in establishing an HTS facility where all of the equipment
needed to support a broad range of HTS applications is readily available. Institutional and NIH
funds (>$4.5 million) have been invested to acquire leading-edge laboratory automation, liquid
handlers, readers and LIMS. These systems include:
 Fully-automated and environmentally-controlled system for compound library management,
storage, retrieval (Nexus/Brooks Universal Store) including a fully-integrated automated
instrumentation suite for production of compound plates for screening (including highcapacity cherry picking and serial dilution).
 Molecular Device ImageXpress Micro XL fully automated microscope imager for high
content screening including a widefield CMOS camera, a wide range of objective lenses and
fluorescent filters as well as phase contrast transmitted light and compatible for 6-1536 well
formats.
 WaveFront Biosciences Panoptic kinetic imaging plate reader. The Panoptic is a second
generation kinetic imaging plate reader that provides increased temporal and spatial
resolution compared to previously developed technologies as well as greater flexibility for
optical and liquid handling settings. Experimental settings provide between 10 excitation and
5 emission wavelengths with 40 ms wavelength switching enabling assays measuring more
than one fluorescent indicator. Its luminescence mode supports assays such as the cAMP
gloSensor, BRET, and SPA. The liquid handling platform allows for 9 nest positions, multiple
tip boxes, plate lidding and delidding, and two-solvent tip washing.
 A Hamamatsu FDSS (Functional Drug Screening System) kinetic imaging plate reader for
running kinetic fluorescence-based and luminescence assays. This instrument has
integrated liquid handling and is capable of simultaneously making liquid additions to each
well of either 384- or 96-well microplates as well as enabling multiple liquid additions to a
test plate during the course of an experiment. The FDSS acquires kinetic data from all wells
of a microtiter plate simultaneously at rates of up to 10 Hz. It is ideal for running the cellbased functional screens of enzymes, ion channels, transporters, and seventransmembrane receptors.
 Perkin Elmer EnSpire and Biotek NEO multimode plate readers. These instruments are
suited to support stable endpoint/slow kinetic assays in absorbance, fluorescence, and
luminescence modes in 96-1536 well formats. Together, these instruments can perform
fluorescence polarization, time-resolved fluorescence, and simultaneous dual emission
fluorescence, luminescence, AlphaScreen and label-free Epic assays. The Biotek NEO is
equipped with a 50-plate capacity stacker.
 Perkin Elmer Topcount plate reader to count radionuclide-based or luminescent samples,
without interfering crosstalk.
 Numerous contact-based liquid handlers to support test compound and reagent addition
including a VPrep (Agilent) and three Bravos (Agilent) for high-precision 96 -1536 well
pipetting, including one in a tissue culture hood for sterile environments.




Numerous liquid handlers to support bulk reagent addition and cell plating in 96-1536 well
plates including six Multidrop Combis (Thermo Fisher) with one contained in a tissue culture
hood
Various other instruments to support the proposed research including two VSpin (Velocity11)
automated plate centrifuges, two Cytomat2 (Kendro) automated cell culture incubators,
PlateLoc (Agilent) plate sealer, two Elx405CW (Bio-Tek) plate washers, XCap tube
decapper (Brooks), and XPeel plate desealer (Brooks).
Two Echo555 (Labcyte) acoustic plate reformatters. These revolutionary, non-contact, ultralow volume (2-1000 nl), random-access liquid handlers are ideal for compound management
and presentation for both cell-based functional assays and for low volume scintillation
proximity assays (SPA).One of these instruments is calibrated for DMSO solutions. The
other is calibrated for an array of solvents allowing the transfer of genomic reagents,
biochemical reagents and buffers.
Seahorse XFe96 Extracellular Flux Analyzer. This device simultaneously interrogates the two
major energy-producing pathways of the cell – mitochondrial respiration and glycolysis. The
Analyzer determines in vitro oxygen consumption rate (OCR) and extracellular acidicfication
rate (ECAR), in order to assess cellular functions such as oxidative phosphorylation,
glycolysis, and fatty acid oxidation.
Three robotically integrated systems support high-content imaging (Molecular Devices
ImageXpress Micro XL and supporting instruments), kinetic imaging plate reading (Hamamatsu
FDSS and supporting instruments), and compound plate creation (Labcyte Echo 555 and
supporting instruments), Plate movement between instruments occurs through a Thermo
Scientific F3 six-axis articulated robotic arm controlled by Thermo Scientific Momentum
scheduling software. This enables walk-away assay execution with time-locked, trackable
events.
All readers on the system are connected via intranet to the facility’s network enabling automated
data acquisition, analysis, visualization, and archival using a variety of commercially acquired
and in-house designed data management and informatics tools including Accelrys Pipeline Pilot,
Accelrys Accord, Microsoft SharePoint, DeltaSoft’s ChemCart suite, and WaveGuide, an inhouse data analysis application. All data are stored in an Oracle database. The facility’s data
management infrastructure will support data deposition into external databases including
PubChem and D360.
Compound Collections:
The Vanderbilt Screening Facility utilizes an ever-growing compound collection designed and
deployed to enable a wide-variety of screening modalities from assay development and pilot
screen to large-scale high-throughput screening (Tables 1 and 2). Our latest addition to our
diverse screening collection includes a 100,000 small molecule library (Life Chemicals) that was
selected by a team of medicinal and computational chemists to ensure diversity and minimize
the number of compounds identified as likely yielding false positive results due to their
containing known reactive groups or scaffolds identified to be promiscuous inhibitors. Our VICB
discovery collection totals ~160,000 compounds; this library was obtained from ChemBridge
and ChemDiv and were chosen to maximally represent the chemical diversity of the larger
compound collections (~1.2 million samples in total). A 77,000 member selection from the
MLPCN compound library is available for screening.
Table 1 VHTS Diverse Screening Collections
NAME
# COMPOUNDS
DESCRIPTION
Vanderbilt Discovery Collection
100,000
Selected from Life Chemicals collection for HTS. The compounds in this collection
were chosen by Vanderbilt medicinal and computational chemists to provide leadlike motifs, minimum pan-assay interference, and maximum diversity.
VICB Collection
160,000
Drug-like, diverse set of small molecules from Chembridge and ChemDiv
Molecular Libraries Small Molecule
Repository (MLSMR)*
77,000
Recent acquisition for MLPCN screens; requires screening results deposition into
Pubchem
Fesik Fragment Library
16,000
Diverse collection of fragment molecules from 8 vendors, rule of 3 compliant, and
filtered against non-specific and interfering molecules.
Other libraries include several collections from the NCI including the approved oncology set, a
51-member epigenetics collection (SelleckChem), 6200-member ion channel library (Life
Chemicals), Spectrum Collection (Microsource), the NIH Clinical Collections I and II (DPI),
Roche Published Kinase Inhibitor Collection, GSK Kinase Inhibitor Collection, Enzo Screen-well
kinase inhibitor library, and an 800-member NCI purified natural products collection. In
partnership with the Vanderbilt Synthesis Core, the size and diversity of the library via internal
and external efforts in both synthetic and technology-enable natural products isolation is
continually growing.
Table 2 VHTS Targeted Collections
NAME
# COMPOUNDS
DESCRIPTION
119
contains most current FDA-approved anticancer drugs. The current set (AOD VI)
consists of 119 agents and is intended to enable cancer research, drug discovery
and combination drug studies.
Diversity Set V
1593
During the generation of the diversity set, the pharmacophores for any candidate
compound are compared to the set of all pharmacophores found in structures
already accepted into the set. If the current structure has more than 5 new
pharmacophores, it is added to the set. An additional objective with the NCI
Diversity Set III was to create a diverse set of compounds that were amenable to
forming structure-based hypotheses.
Mechanistic Set III
813
was derived from the 37,836 open compounds that have been tested in the NCI
human tumor 60 cell line screen. In contrast to the original diversity set of 1,990
compounds, which was chosen on the basis of structural diversity
Natural Product Set III
117
This set was created in response to numerous drug discovery research groups
that expressed a desired to study a variety of scaffold structures having multiple
functional groups.
NCI Focused Natural Product Collection
819
Pure compounds acquired by the NCI from Analytical and MerLion
Approved Oncology Drugs Set VI
Spectrum Collection
NIH Clinical Collection I and II
Cayman Lipid Library
Marnett Collection
Ion Channel
Epigenetics Collection
Enzo Kinase Inhibitor Library
2,000
730
1,000
215
>6000
A wide range of biologically active and structurally diverse compounds. 50% drug
components, 30% natural products, 20% other bioactive components.
Small molecules that have history of use in human clinical trials.
Broad variety of bioactive lipids.
NSAID derivatives that contain cyclooxygenase inhibitors, PPARgamma activators
and apoptosis inducers.
A collection from Life Chemicals targeted to ion channels compiled using 2D
fingerprint similarity methodology.
51
A group of small molecule modulators with biological activity for use in epigenetic
research.
80
The Screen-Well™ Kinase Inhibitor Library contains 80 known kinase inhibitors of
well-defined activity. Includes inhibitors of these important kinases: Insulin/IGF
Receptors, PI 3-Kinase, CaM Kinase II, JAK, PKA, CDK, JNK, PKC, CKI II, MAPK,
RAF, EGFR, MEK, SAPK, GSK, MLCK, Src-family, IKK, PDGFR, VEGFR, and many
more.
GSK Published Kinase Inhibitor Library*
349
A set of 349 small molecule kinase inhibitors spanning over 30 chemotypes that
have been previously published by GlaxoSmithKline in peer-reviewed scientific
journals.
Roche Published Protein Kinase Inhibitor
Library*
235
Small molecule protein kinase inhibitors disclosed in peer-reviewed scientific
publications.
* requires collaborative agreement with supplier
siRNA Libraries:
In 2013, the HTS Facility expanded screening capabilities to the genomic level by adding
several human siRNA libraries (Dharmacon). These are available in 96 or 384-well plates in a
ready-to-transfect format. Included in this collection are pooled kinase, individual kinase, pooled
druggable genome, and whole-genome libraries.
Personnel:
Paige Vinson, Ph.D.: Director and Research Assistant Professor of Biochemistry
C. David Weaver, Ph.D.: Scientific Director and Associate Professor of Pharmacology
Joshua Bauer, Ph.D.: Director of High-content Imaging and Functional Genomic Screening,
Research Assistant Professor of Biochemistry
Chris Farmer, Ph.D.: Director of Screening Informatics
Emily Days: Drug Discovery Scientist
Debbie Mi, Ph.D.: Drug Discovery Scientist
Corbin Whitwell: Compound Management Specialist
Erin Gribben: Scientific Research Core Facility Coordinator
Mike McCaughey, Ph.D.: Automation Engineer, Research Assistant Professor
Lisa Wright: Core Manager
Matthew Graves: Software Programmer
David Baughman: Computer Systems Administrator