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High Throughput Screening Facility
The University of Iowa High Throughput Screening Facility (HTS) was established in 2011 through the generous support of a collection of units on campus, including: the College of Pharmacy, Carver College of Medicine, Departments of Biochemistry, Pharmacology, Anatomy & Cell Biology, Physiology, Microbiology, Internal Medicine, the Holden Comprehensive Cancer Center and the Institute for Clinical and Translational Science. The UI HTS Facility operates as an Office of The Vice President For Research Core Facility.
Current Capability and Instrumentation Summary
 Located in Room 310 in the College of Pharmacy, The University of Iowa High Throughput Screening Facility (UIHTS) is currently equipped to perform screening of small molecule libraries in 96, 384 and 1536-well format using a robotic liquid handler integrated with plate reader detection.
Robotics: Our current robotic platform is a Hamilton Star liquid handler equipped with 8-independent channels and a 384 channel pipetting head. The liquid handling robot is also equipped with on-board incubators (4 plates) and two positions for incubation/chilling of stock solutions. The platform also includes a tip washer for consumable savings when needed.
Detection: Integrated with the Hamilton Star is a Perkin Elmer Envision Multimode high speed plate reader equipped with filters and monochromators. The plate reader, controlled through the integrated Hamilton software, supports fluorescence, time-resolved fluorescence, fluorescence polarization, time-resolved FRET, absorbance and specialized screening technologies AlphaScreen, DELFIA, and LANCE. The Envision is equipped with two photomultipliers and a dedicated uHTS laser for high speed AlphaScreen. Typical read times on the plate reader range from 45s to 6 minutes, depending on well density. High speed “on the fly” measurements can allow for reading a 384 well plate in as little as 36 seconds.
The laboratory is also equipped with a 5-color Roche Lightcycler 480 II system, which allows for secondary confirmation of hits binding to a purified target using Differential Scanning Fluorimetry, also known as Thermal-melt analysis.
 Libraries: UIHTS is currently stocked with two small molecule libraries. The Spectrum Library from MicroSource consists of 2320 structurally diverse compounds including marketed and experimental drugs as well as natural products. This array of test compounds provides a rigorous platform for assay validation and testing. The larger collection is from Chembridge, the Diverset, a collection of 50,000 small molecules representing a wide swath of chemical space, optimized to be “drug like,” considering factors such as partition coefficient and Lipinski-like rules.
Data Management: Management of compound library plates and screening data is done through M-Screen, a web-accessed academic database solution for High Throughput Screening data analysis and curation.
The UIHTS lab is also equipped with other support instrumentation, such as a plate centrifuge, water polisher, and automated thermal plate sealer.
High Throughput Screening Facility - Phase II (Spring-Summer 2012)
Due to the success of an NIH High End Equipment proposal (1S10RR029274), the University of Iowa is in the process of enacting Phase II of our planned High Throughput Screening Facility. Phase II will significantly expand the capabilities of the center to allow for the use of live cells and high content imaging.
The core technology of Phase II is the Perkin Elmer Cell::Explorer robotic system completed contained within a custom HEPA-filtered enclosure. The system has a six-axis industrial robotic arm, which can access and move plates and labware between the following integrated equipment and instruments:
- Janus Mini MDT liquid handler, for dispensing and transferring liquids in 96, 384 and 1536-well plates.
- Liconic LPX220 Plate hotel, for storage of up to 44 plates and 40 tip boxes
- Liconic STX44 CO2 humidified cell incubator with 44 plate capacity
- BioTek Elx405UCW bulk liquid dispenser and plate washer
- FlexDrop III, 2 channel bulk liquid dispenser
- Operetta High Content Imaging System
The detection modality of the cell::explorer system high content, or live cell imaging, by the Operetta High Content system. The Operetta is a true confocal high throughput microscopic imaging system, that includes software for high speed image analysis for quantifying measurable cellular phenomena, such as changes in subcellular localization of a tagged target protein, changes in cell membrane shape or composition, as well as colocalization quantification. The system can essentially perform any experiment one could do with a standalone confocal microscope, but in a massively parallel high throughput fashion. The automated, centralized control of the system also allows collection of images as discrete timepoints (minutes, hours, days, weeks) with automated robotic control of plate retrieval from the incubator, to imaging and analysis on the Operetta.
Perhaps most important is the ability of the unified system to examine the effect of individual compounds from our small molecule libraries on an investigator’s cellular process or target of interest. The Operetta control software includes a large library of predefined cellular phenomena measurements, as well as libraries of customizable measurement functions, and the ability for advanced users to write their own image analysis scripts.
In addition to customized assays for drug discovery applications, a wide array of vendors offer turn-key kits for the measurement of cellular phenomena, including apoptosis, autophagy (LC3B & Poly-ubiquitin), caspase activation (3 & 9), cell viability, PARP cleavage, cytochrome C detection, mitosis and many others.
The small molecule libraries currently housed in the High Throughput Screening facility can be used with this cellular HCS system, however, the system is very powerful, and could be used for RNAi library screening as well. Currently, the facility does not house an RNAi library.
Additional technical information on the Operetta’s capabilities can be found on the vendor’s website:
http://www.perkinelmer.com/pages/020/cellularimaging/products/operetta.xhtml
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