Hardware Capability (Clink for throughput capability and target applications in ChemCORE)

GENERAL DESCRIPTION

ChemCORE houses several liquid handlers including one Tekbench™ Work Station, two Cybi-Well™ systems, and BioMek2000™ work station. They are capable of handling 96- and 384-well plates in a variety of formats including high throughput liquid handling, cherry-picking and volume dispensing. The detection modules include the BD Pathway™ 855 Bioimager, Epic ™ System, Tecan Safire 2 reader, Aquest™ (currently Analyst GT) plate reader, ICR-8000™ atomic absorption spectrometer, SpectraMax™ 340 reader, and LAS-3000 Fuji imaging station. The liquid handling and detection module are highly integrated by a Mitsubishi RV-2AJ robotic arm and Zymark Twister™ II arm. In addition, both liquid handling modules and detection modules are robotically linked to accessory units including a Kendro Cytomat 6070 automated incubator, Elx-405 plate washers, and Multidrop dispensers.

The integration and performance of liquid handlers, detection modules and accessory equipment are controlled by customized a Genera™ software package, which allows for multi-tasking of instrument clusters, web-based remote monitor/control, as well as stand-alone performance of individual units. The data acquisition, analyses, and storage are carried out by several CPUs and managed by an Oracle server along with several software packages to perform SAR and chemoinformatics analyses. Data access and analyses can be performed within our ChemCORE-maintained network.

DETECTION INSTRUMENTS

FDSS6000 System, a high-throughput kinetic fluorescence reader by Hamamatsu

http://www.fdssdrug.com

 

Brief description  ( for throughput details )

The FDSS6000 System is an imaging based plate reader for cellular assays, assay development and high throughput screening which can utilize both fluorescence and luminescence modalities. FDSS 6000 can respond to various fluorescent probes, and can perform cell-based assay at high speed and with high precision. Since it corresponded to High Throughput Screening (HTS), the multi-dispenser and the micro plate automatic handling system were equipped, and improvement in the speed and automation were realized. The fluorescence observation application of ion imaging and FRET can be supported, and the dispenser of a maximum of 3 sets can be loaded. Simultaneous loading of the object for fluorescence and the camera for luminescence is possible.

 

Uses and applications

 

1. Netzer, R., Bischoff, U. & Ebneth, A. HTS techniques to investigate the potential effects of compounds on cardiac ion channels at early-stages of drug discovery. Current Opinion in Drug Discovery & Development 6, 462 - 469 (2003).

 

2. Kim, H. S. et al. Design, synthesis, and biological evaluation of 1,3-dioxoisoindoline-5-carboxamide derivatives as T-type calcium channel blockers. Bioorganic & Medicinal Chemistry Letters 17, 476-481 (2007).

 

3. Poul, E. L. et al. Adaptation of Aequorin Functional Assay to High Throughput Screening. J Biomol Screen 7, 57-65 (2002).

 

Epic ™ System, a high-throughput label-free detection platform from Corning®

http://www.corning.com

 

Brief description  ( for throughput details )

 

The Epic™ System is a new high-throughput label-free detection platform from Corning, Inc, consisted of an SBS-standard 384-well microplate with optical sensors and HTS-compatible microplate reader capable of reading up to 40,000 wells per 8 hours and a set of label-free, direct-bind and functional assay protocols. Its sensitivity of 5pg/mm2 enables the detection of the binding of a 300Da compound to a 70kDa immobilized target with CVs of 10% or less. This powerful combination of features and performance capabilities not only enables researchers to evaluate many of the biomolecular interactions in molecular and cellular biology, but also provides the added benefit of integration with an already installed HTS capital base. In addition, the Epic™ System makes the screening of "intractable" targets and pathway interactions that cannot be screened today possible. The universal applicability of the Epic™ System across biochemical assay types also enables the discovery of new chemical entities (small molecule) as well as new biological entities (large molecule).

 

Uses and applications

 

1. Wu, Meng; Coblitz, Brian; Shikano, Sojin; Long, Shunyou; Spieker, Matt; Frutos, Anthony G.; Mukhopadhyay, Sunil; Li, Min, Phospho-specific recognition by 14-3-3 proteins and antibodies monitored by a high throughput label-free optical biosensor. FEBS Letters, In Press.

 

2. Fang, Ye; Ferrie, Ann M.; Fontaine, Norman H.; Mauro, John; Balakrishnan, Jitendra, Resonant Waveguide Grating Biosensor for Living Cell Sensing. Biophys. J. 2006, 91, (5), 1925-1940.

 

3. Fang, Ye; Ferrie, Ann M.; Li, Guangshan, Cellular functions of cholesterol probed with optical biosensors. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research 2006, 1763, (2), 254-261.

 

Ion Channel Reader 8000 and 12000, an atomic absorption spectroscopy and flux assay based ion channel activity detection system from Aurora Biomed Inc.

 

http://www.aurorabiomed.com

 

 

Brief description  ( for throughput details )

 

The ICR 8000 detects ion channel activity using atomic absorption spectroscopy and flux assays. Fux assays utilize tracer ions to study the ion channel of interest.  The Ion Channel Reader 8000 provides a medium throughput format up to 5000 data points/day for ion channel screening by the flux assay approach.  The ICR 8000 displays broad utility; it can be applied to assess voltage-gated (hERG, BK/SK, Kv1.1, 1.4, 1.5, KCNQ, 2P, etc.) and ligand-gated (KATP, nAChR, etc.) potassium channels, voltage-gated sodium channels (SCN5a, Nav1.2) and transporters (Na/K-ATPase, K-Cl Co-transporter). This allows researchers to accelerate drug development efforts in the treatment and prevention of diseases associated with ion channels. It can accommodate both 96 and 384 well plates with CV less than 5%.

 

The ICR 12000 system is essentially identical to ICR8000.  the primary difference lies that it is a 12 burners instead of a single burner in ICR8000.  Hence, ICR12000 increases the throughput by a factor of 12.

 

Uses and applications

 

 

1. Terstappen, Georg C., Functional Analysis of Native and Recombinant Ion Channels Using a High-Capacity Nonradioactive Rubidium Efflux Assay. Analytical Biochemistry 1999, 272, (2), 149-155.
 

2. Tang, Weimin; Kang, Jiesheng; Wu, Xiaying; Rampe, David; Wang, Lin; Shen, Hong; Li, Zhuyin; Dunnington, Damien; Garyantes, Tina, Development and Evaluation of High Throughput Functional Assay Methods for hERG Potassium Channel. J Biomol Screen 2001, 6, (5), 325-331.

 

3. Terstappen, Georg C., Nonradioactive Rubidium Ion Efflux Assay and Its Applications in Drug Discovery and Development. ASSAY and Drug Development Technologies 2004, 2, (5), 553-559.

 

4. Sun H, Shikano S, Xiong Q & Li M (2004). Function recovery after chemobleaching (FRAC): evidence for activity silent membrane receptors on cell surface. Proc Natl Acad Sci USA 101(48):16964-9.

 

5. Sun, Haiyan; Liu, Xiaodong; Xiong, Qiaojie; Shikano, Sojin; Li, Min, Chronic Inhibition of Cardiac Kir2.1 and hERG Potassium Channels by Celastrol with Dual Effects on Both Ion Conductivity and Protein Trafficking. J. Biol. Chem. 2006, 281, (9), 5877-5884.

 

BD Pathway™ 855 Bioimager, an automated, confocal, real-time, single cell kinetic and endpoint imaging system from BD Biosciences.

http://www.bdbiosciences.com

 

Brief description  ( for throughput details )

 

The BD Pathway Bioimager is an automated, confocal, real-time, single cell kinetic and endpoint imaging system that has been integrated into a single, compact unit.  The BD Pathway was designed to provide high-resolution, automated confocal imaging with sophisticated imaging software to assist in developing better cell-based assays. The system allows the user to explore biological events without many of the restrictions of conventional microscope-based high content imaging systems.

BD Pathway offers:

§         True confocal real-time imaging

§         Full-spectrum laser-free illumination

§         Ability to run kinetic and endpoint assay formats

§         Integrated liquid handling with image-as-you-add capability

§         96- and 384-multiwell plate and slide imaging

§         Environmentally controlled for live cell experiments

§         Flexible software allowing easy data navigation and classification

§         Integrated binocular viewing

§         Ultra-high precision linear x,y, and z motors

 

Uses and applications

 

1. Chan, Grace K. Y.; Richards, Gillian R.; Peters, Marco; Simpson, Peter B., High Content Kinetic Assays of Neuronal Signaling Implemented on BDTM Pathway HT. ASSAY and Drug Development Technologies 2005, 3, (6), 623-636.

 

2. Lang, Paul; Yeow, Karen; Nichols, Anthony; Scheer, Alexander, Cellular imaging in drug discovery. Nat Rev Drug Discov 2006, 5, (4), 343-356.

 

3. Timothy, J. Mitchison, Small-Molecule Screening and Profiling by Using Automated Microscopy. ChemBioChem 2005, 6, (1), 33-39

 

 

Safire²™ plate reader, a monochrometer based multifuntional plate reader from Tecan.

 

http://www.tecan.com

 

Brief description  ( for throughput details )

 

Safire² has been developed with the drug discovery process in mind. Due to its in-built flexibility, sensitivity and performance, Safire² is the ideal instrument to bridge the gap between assay development and high throughput screening. Safire² is a fully modular monochromator-based microplate detection system that offers a range of high speed detection techniques. In assay development, which requires frequent changing of applications and fluorophores, the monochromator-based Safire² eliminates the need for cumbersome filter changes thereby saving time and money.

The modular concept of Safire² allows upgrades to new detection modes at any time if there is a need to have access to further applications. Detection modules are available for top and bottom fluorescence intensity measurements, fluorescence polarization studies and multi-channel absorbance and luminescence measurements. Safire² is easily combined with a stacker module for batch processing of up to 50 microplates.

 

Uses and applications

 

1. Park SH, Raines RT. Fluorescence polarization assay to quantify protein-protein interactions. Methods Mol Biol. 2004;261:161-6.


2. Lakowicz, J.R.  Principles of Fluorescence Spectroscopy, 2nd ed., Plenum, New York, 1999.

 

3. Valeur, B. Molecular Fluorescence, Wiley-VCH, Weinheim, 2001.

 

4. Wu, Meng; Coblitz, Brian; Shikano, Sojin; Long, Shunyou; Cockrell, Lisa M.; Fu, Haian; Li, Min, SWTY-A general peptide probe for homogeneous solution binding assay of 14-3-3 proteins. Analytical Biochemistry 2006, 349, (2), 186-196.

 

5. Coblitz, Brian; Shikano, Sojin; Wu, Meng; Gabelli, Sandra B.; Cockrell, Lisa M.; Spieker, Matt; Hanyu, Yoshiro; Fu, Haian; Amzel, L. Mario; Li, Min, C-terminal Recognition by 14-3-3 Proteins for Surface Expression of Membrane Receptors. J. Biol. Chem. 2005, 280, (43), 36263-36272.

 

 

Aquest™ (currently Analyst GT) plate reader, a filter based multifunction plate reader from LJL system (currently Molecular Devices).

 

http://www.moleculardevices.com

 

 

Brief description  ( for throughput details )

 

Analyst set the standard for no-compromise performance in 384-well plates when it was introduced. Acquest, the first commercially available 1536 multimode reader, brought that performance to high-density microplates. Now Analyst GT builds on that reputation with significantly faster read speeds for all plate formats. Analyst GT achieves this performance with second-generation SmartOptics™, a unique system that selects and configures light sources, filters, detectors, and optical paths to ensure maximum assay performance. Previously, systems that offered 96- through 1536-well compatibility sacrificed read time or sensitivity to read high-density 1536-well plates. Analyst GT improves on Acquest, the benchmark for 1536-well point reading systems, by dramatically increasing read speed without compromising signal-to-noise and signal-to-background.

 

Uses and applications

 

1. Park SH, Raines RT. Fluorescence polarization assay to quantify protein-protein interactions. Methods Mol Biol. 2004;261:161-6.


2. Lakowicz, J.R.  Principles of Fluorescence Spectroscopy, 2nd ed., Plenum, New York, 1999.

 

3. Valeur, B. Molecular Fluorescence, Wiley-VCH, Weinheim, 2001

 

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