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featuring the Echo® Acoustic Technology
PUBLICATIONS

by scientists and researchers like you

103 Total Publications
HOME | RESOURCES | Publications

PUBLICATIONS

103 Total Publications

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TITLES and AUTHORS Year Link PDF + Abstract
  • High-Throughput Screening Using iPSC-Derived Neuronal Progenitors to Identify Compounds Counteracting Epigenetic Gene Silencing in Fragile X Syndrome Kaufmann M, et al.

    Institution: Center of Proteomic Chemistry, Novartis Institutes for Biomedical Research, Novartis Campus, Basel

    Publication: J Biomol Screen

    2015 abstract

    Fragile X syndrome (FXS) is the most common form of inherited mental retardation, and it is caused in most of cases by epigenetic silencing of the Fmr1 gene. Today, no specific therapy exists for FXS, and current treatments are only directed to improve behavioral symptoms. Neuronal progenitors derived from FXS patient induced pluripotent stem cells (iPSCs) represent a unique model to study the disease and develop assays for large-scale drug discovery screens since they conserve the Fmr1 gene silenced within the disease context. We have established a high-content imaging assay to run a large-scale phenotypic screen aimed to identify compounds that reactivate the silenced Fmr1 gene. A set of 50,000 compounds was tested, including modulators of several epigenetic targets. We describe an integrated drug discovery model comprising iPSC generation, culture scale-up, and quality control and screening with a very sensitive high-content imaging assay assisted by single-cell image analysis and multiparametric data analysis based on machine learning algorithms. The screening identified several compounds that induced a weak expression of fragile X mental retardation protein (FMRP) and thus sets the basis for further large-scale screens to find candidate drugs or targets tackling the underlying mechanism of FXS with potential for therapeutic intervention.

    Publication / Type:
    J Biomol Screen
    Related Subject:
    high-content screening, fragile X syndrome, induced pluripotent stem cells
    Link:
    http://jbx.sagepub.com/content/20/9/1101.abstract
  • Utilizing Low-Volume Aqueous Acoustic Transfer with the Echo 525 to Enable Miniaturization of qRT-PCR Assay S Agrawal, S Cifelli, R Johnstone, D Pechter, D Barbey, K Lin, T Allison, S Agrawal, A Rivera-Gines,

    Institution: Merck Research Laboratories; Labcyte Inc; Case Western Reserve University School of Medicine

    Publication: Journal of Laboratory Automation (JALA) Special Issue

    2015 abstract

    Quantitative reverse transcription PCR (qRT-PCR) is a valuable tool for characterizing the effects of inhibitors on viral replication. The amplification of target viral genes through the use of specifically designed fluorescent probes and primers provides a reliable method for quantifying RNA. Due to reagent costs, use of these assays for compound evaluation is limited. Until recently, the inability to accurately dispense low volumes of qRT-PCR assay reagents precluded the routine use of this PCR assay for compound evaluation in drug discovery. Acoustic dispensing has become an integral part of drug discovery during the past decade; however, acoustic transfer of microliter volumes of aqueous reagents was time consuming. The Labcyte Echo 525 liquid handler was designed to enable rapid aqueous transfers. We compared the accuracy and precision of a qPCR assay using the Labcyte Echo 525 to those of the BioMek FX, a traditional liquid handler, with the goal of reducing the volume and cost of the assay. The data show that the Echo 525 provides higher accuracy and precision compared to the current process using a traditional liquid handler. Comparable data for assay volumes from 500 nL to 12 µL allowed the miniaturization of the assay, resulting in significant cost savings of drug discovery and process streamlining.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    acoustic droplet ejection, HTS, high-throughput screening, automated biology, molecular biology, automated biology, genomics, drug discovery
    Link:
    http://jla.sagepub.com/content/21/1/57.full.pdf+html
  • Miniaturization of High-Throughput Epigenetic Methyltransferase Assays with Acoustic Liquid Handling Bonnie Edwards, John Lesnick, Jing Wang, Nga Tang, Carl Peters

    Institution: Labcyte Inc; Cisbio Bioassays; BMG Labtech

    Publication: Journal of Laboratory Automation (JALA) Special Issue

    2015 abstract

    Epigenetics continues to emerge as an important target class for drug discovery and cancer research. As programs scale to evaluate many new targets related to epigenetic expression, new tools and techniques are required to enable efficient and reproducible high-throughput epigenetic screening. Assay miniaturization increases screening throughput and reduces operating costs. Echo liquid handlers can transfer compounds, samples, reagents, and beads in submicroliter volumes to high-density assay formats using only acoustic energy—no contact or tips required. This eliminates tip costs and reduces the risk of reagent carryover. In this study, we demonstrate the miniaturization of a methyltransferase assay using Echo liquid handlers and two different assay technologies: AlphaLISA from PerkinElmer and EPIgeneous HTRF from Cisbio.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    epigenetics, methyltransferase, ADE (acoustic droplet ejection), liquid handling, HTRF, AlphaLISA
    Link:
    http://jla.sagepub.com/content/21/1/208.full.pdf+html
  • The Evolution of MALDI-TOF Mass Spectrometry toward Ultra-High- Throughput Screening: 1536-Well Format and Beyond Carl Haslam, et al.

    Institution: Department of Biological Sciences, GlaxoSmithKline

    Publication: J Biomol Screen

    2015 abstract

    Mass spectrometry (MS) offers a label-free, direct-detection method, in contrast to fluorescent or colorimetric methodologies. Over recent years, solid-phase extraction–based techniques, such as the Agilent RapidFire system, have emerged that are capable of analyzing samples in <10 s. While dramatically faster than liquid chromatography–coupled MS, an analysis time of 8–10 s is still considered relatively slow for full-diversity high-throughput screening (HTS). Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) offers an alternative for high-throughput MS detection. However, sample preparation and deposition onto the MALDI target, as well as interference from matrix ions, have been considered limitations for the use of MALDI for screening assays. Here we describe the development and validation of assays for both small-molecule and peptide analytes using MALDI-TOF coupled with nanoliter liquid handling. Using the JMJD2c histone demethylase and acetylcholinesterase as model systems, we have generated robust data in a 1536 format and also increased sample deposition to 6144 samples per target. Using these methods, we demonstrate that this technology can deliver fast sample analysis time with low sample volume, and data comparable to that of current RapidFire assays.

    Publication / Type:
    J Biomol Screen
    Related Subject:
    MALDI-TOF, mass spectrometry, high-throughput screening, RapidFire
    Link:
    http://jbx.sagepub.com/content/early/2015/10/01/1087057115608605.abstract
  • Technologies That Enable Accurate and Precise Nano- to Milliliter-Scale Liquid Dispensing of Aqueous Reagents Using Acoustic Droplet Ejection E Sackmann, L Majlof1, A Hahn-Windgassen, B Eaton, T Bandzava, J Daulton, A Vandenbroucke, M Mock, R

    Institution: Labcyte Inc.

    Publication: Journal of Laboratory Automation (JALA) Special Issue

    2015 abstract

    Acoustic liquid handling uses high-frequency acoustic signals that are focused on the surface of a fluid to eject droplets with high accuracy and precision for various life science applications. Here we present a multiwell source plate, the Echo Qualified Reservoir (ER), which can acoustically transfer over 2.5 mL of fluid per well in 25-nL increments using an Echo 525 liquid handler. We demonstrate two Labcyte technologies—Dynamic Fluid Analysis (DFA) methods and a high-voltage (HV) grid—that are required to maintain accurate and precise fluid transfers from the ER at this volume scale. DFA methods were employed to dynamically assess the energy requirements of the fluid and adjust the acoustic ejection parameters to maintain a constant velocity droplet. Furthermore, we demonstrate that the HV grid enhances droplet velocity and coalescence at the destination plate. These technologies enabled 5-µL per destination well transfers to a 384-well plate, with accuracy and precision values better than 4%. Last, we used the ER and Echo 525 liquid handler to perform a quantitative polymerase chain reaction (qPCR) assay to demonstrate an application that benefits from the flexibility and larger volume capabilities of the ER.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    acoustic droplet ejection, reservoir, droplet velocity, droplet bounce, aqueous
    Link:
    http://jla.sagepub.com/content/21/1/166.full.pdf+html
  • Mammalian Genotyping Using Acoustic Droplet Ejection for Enhanced Data Reproducibility, Superior Throughput, and Minimized Cross-Contamination C Cain-Hom, R Pabalate, A Pham, H Patel, R Wiler, J Colin Cox

    Institution: Department of Transgenic Technology, Genentech Inc.

    Publication: Journal of Laboratory Automation

    2015 abstract

    Genetically engineered animal models are major tools of a drug discovery pipeline because they facilitate understanding of the molecular and biochemical basis of disease. These highly complex models of human disease often require increasingly convoluted genetic analysis. With growing needs for throughput and consistency, we find that traditional aspiration-and-dispense liquid-handling robots no longer have the required speed, quality, or reproducibility.

    We present an adaptation and installation of an acoustic droplet ejection (ADE) liquid-handling system for ultra-high-throughput screening of genetically engineered models. An ADE system is fully integrated with existing laboratory processes and platforms to facilitate execution of PCR and quantitative PCR (qPCR) reactions. Such a configuration permits interrogation of highly complex genetic models in a variety of backgrounds. Our findings demonstrate that a single ADE system replaces 8–10 traditional liquid-handling robots while increasing quality and reproducibility.

    We demonstrate significant improvements achieved by transitioning to an ADE device: extremely low detectable cross-contamination in PCR and qPCR despite extensive use, greatly increased data reproducibility (large increases in data quality and Cq consistency), lowered reaction volumes for large cost savings, and nearly a magnitude increase in speed per instrument. We show several comparisons between traditional- and ADE-based pipetting for a qPCR-based workflow.

    Publication / Type:
    Journal of Laboratory Automation
    Related Subject:
    genotyping
    Link:
    http://jla.sagepub.com/content/early/2015/08/26/2211068215601637.abstract
  • Mammalian Genotyping Using Acoustic Droplet Ejection for Enhanced Data Reproducibility, Superior Throughput, and Minimized Cross-Contamination C Cain-Hom, R Pabalate, A Pham, H Patel, R Wiler, J Cox

    Institution: Department of Transgenic Technology, Genentech

    Publication: Journal of Laboratory Automation (JALA) Special Issue

    2015 abstract

    Genetically engineered animal models are major tools of a drug discovery pipeline because they facilitate understanding of the molecular and biochemical basis of disease. These highly complex models of human disease often require increasingly convoluted genetic analysis. With growing needs for throughput and consistency, we find that traditional aspiration-and-dispense liquid-handling robots no longer have the required speed, quality, or reproducibility. We present an adaptation and installation of an acoustic droplet ejection (ADE) liquid-handling system for ultra-high-throughput screening of genetically engineered models. An ADE system is fully integrated with existing laboratory processes and platforms to facilitate execution of PCR and quantitative PCR (qPCR) reactions. Such a configuration permits interrogation of highly complex genetic models in a variety of backgrounds. Our findings demonstrate that a single ADE system replaces 8–10 traditional liquid-handling robots while increasing quality and reproducibility. We demonstrate significant improvements achieved by transitioning to an ADE device: extremely low detectable cross-contamination in PCR and qPCR despite extensive use, greatly increased data reproducibility (large increases in data quality and Cq consistency), lowered reaction volumes for large cost savings, and nearly a magnitude increase in speed per instrument. We show several comparisons between traditional- and ADE-based pipetting for a qPCR-based workflow.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    genotyping, acoustic droplet ejection, RT-PCR, Echo, ADE
    Link:
    http://jla.sagepub.com/content/21/1/37.full.pdf+html
  • Developments in the Implementation of Acoustic Droplet Ejection for Protein Crystallography P Wu, C Noland, M Ultsch, B Edwards, D Harris, R Mayer, S Harris

    Institution: Department of Structural Biology, Genentech; Labcyte Inc.

    Publication: Journal of Laboratory Automation (JALA) Special Issue

    2015 abstract

    Acoustic droplet ejection (ADE) enables crystallization experiments at the low-nanoliter scale, resulting in rapid vapor diffusion equilibration dynamics and efficient reagent usage in the empirical discovery of structure-enabling protein crystallization conditions. We extend our validation of this technology applied to the diverse physicochemical property space of aqueous crystallization reagents where dynamic fluid analysis coupled to ADE aids in accurate and precise dispensations. Addition of crystallization seed stocks, chemical additives, or small-molecule ligands effectively modulates crystallization, and we here provide examples in optimization of crystal morphology and diffraction quality by the acoustic delivery of ultra-small volumes of these cofactors. Additional applications are discussed, including set up of in situ proteolysis and alternate geometries of crystallization that leverage the small scale afforded by acoustic delivery. Finally, we describe parameters of a system of automation in which the acoustic liquid handler is integrated with a robotic arm, plate centrifuge, peeler, sealer, and stacks, which allows unattended high-throughput crystallization experimentation.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    nanoliter crystallization, acoustic droplet ejection, automation, miniaturization, structural biology, automated biology
    Link:
    http://jla.sagepub.com/content/21/1/97.full.pdf+html
  • Ex Vivo Assessment of Drug Activity in Patient Tumor Cells as a Basis for Tailored Cancer Therapy K Blom, P Nygren, J Alvarsson, R Larsson, C Andersson

    Institution: Department of Medical Sciences, Uppsala University;

    Publication: Journal of Laboratory Automation (JALA) Special Issue

    2015 PDF abstract

    Although medical cancer treatment has improved during the past decades, it is difficult to choose between several first-line treatments supposed to be equally active in the diagnostic group. It is even more difficult to select a treatment after the standard protocols have failed. Any guidance for selection of the most effective treatment is valuable at these critical stages. We describe the principles and procedures for ex vivo assessment of drug activity in tumor cells from patients as a basis for tailored cancer treatment. Patient tumor cells are assayed for cytotoxicity with a panel of drugs. Acoustic drug dispensing provides great flexibility in the selection of drugs for testing; currently, up to 80 compounds and/or combinations thereof may be tested for each patient. Drug response predictions are obtained by classification using an empirical model based on historical responses for the diagnosis. The laboratory workflow is supported by an integrated system that enables rapid analysis and automatic generation of the clinical referral response.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    tumor cell, ex vivo, chemotherapy, tailored cancer therapy
    Link:
    http://jla.sagepub.com/content/21/1/178.full.pdf+html
  • Acoustic Sample Deposition MALDI-MS (ASD-MALDI-MS): A Novel Process Flow for Quality Control Screening of Compound Libraries J Chin, E Wood, G Peters, D Drexler

    Institution: Bristol-Myers Squibb; Department of Molecular Biophysics and Biochemistry, Yale University

    Publication: Journal of Laboratory Automation (JALA) Special Issue

    2015 abstract

    In the early stages of drug discovery, high-throughput screening (HTS) of compound libraries against pharmaceutical targets is a common method to identify potential lead molecules. For these HTS campaigns to be efficient and successful, continuous quality control of the compound collection is necessary and crucial. However, the large number of compound samples and the limited sample amount pose unique challenges. Presented here is a proof-of-concept study for a novel process flow for the quality control screening of small-molecule compound libraries that consumes only minimal amounts of samples and affords compound-specific molecular data. This process employs an acoustic sample deposition (ASD) technique for the offline sample preparation by depositing nanoliter volumes in an array format onto microscope glass slides followed by matrix-assisted laser desorption/ionization mass spectrometric (MALDI-MS) analysis. An initial study of a 384-compound array employing the ASD-MALDI-MS workflow resulted in a 75% first-pass positive identification rate with an analysis time of <1 s per sample.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    acoustic sample deposition, MALDI-MS, high-throughput screening
    Link:
    http://jla.sagepub.com/content/21/1/204.full.pdf+html
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