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featuring the Echo® Acoustic Technology
PUBLICATIONS

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103 Total Publications
HOME | RESOURCES | Publications

PUBLICATIONS

103 Total Publications

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TITLES and AUTHORS Year Link PDF + Abstract
  • Gentle, fast and effective crystal soaking by acoustic dispensing P.M. Collins et. al.

    Institution: Diamond Light Source, Harwell Science and Innovation Campus

    Publication: Acta Crystallographica

    2017 abstract

    The steady expansion in the capacity of modern beamlines for high-throughput data collection, enabled by increasing X-ray brightness, capacity of robotics and detector speeds, has pushed the bottleneck upstream towards sample preparation. Even in ligand-binding studies using crystal soaking, the experiment best able to exploit beamline capacity, a primary limitation is the need for gentle and nontrivial soaking regimens such as stepwise concentration increases, even for robust and well characterized crystals. Here, the use of acoustic droplet ejection for the soaking of protein crystals with small molecules is described, and it is shown that it is both gentle on crystals and allows very high throughput, with 1000 unique soaks easily performed in under 10 min. In addition to having very low compound consumption (tens of nanolitres per sample), the positional precision of acoustic droplet ejection enables the targeted placement of the compound/solvent away from crystals and towards drop edges, allowing gradual diffusion of solvent across the drop. This ensures both an improvement in the reproducibility of X-ray diffraction and increased solvent tolerance of the crystals, thus enabling higher effective compound-soaking concentrations. The technique is detailed here with examples from the protein target JMJD2D, a histone lysine demethylase with roles in cancer and the focus of active structure-based drug-design efforts.

    Publication / Type:
    Acta Crystallographica
    Related Subject:
    acoustic dispensing, high-throughput data, protein crystallography
    Link:
    http://scripts.iucr.org/cgi-bin/paper?S205979831700331X
  • Interlaboratory Reproducibility of Droplet Digital Polymerase Chain Reaction Using a New DNA Reference Material Format Leonardo B. Pinheiro, et. al.

    Institution: National Measurement Institute (NMI), Sydney, Australia, et al.

    Publication: Analytical Chemistry

    2017 abstract

    Use of droplet digital PCR technology (ddPCR) is expanding rapidly in the diversity of applications and number of users around the world. Access to relatively simple and affordable commercial ddPCR technology has attracted wide interest in use of this technology as a molecular diagnostic tool. For ddPCR to effectively transition to a molecular diagnostic setting requires processes for method validation and verification and demonstration of reproducible instrument performance. In this study, we describe the development and characterization of a DNA reference material (NMI NA008 High GC reference material) comprising a challenging methylated GC-rich DNA template under a novel 96-well microplate format. A scalable process using high precision acoustic dispensing technology was validated to produce the DNA reference material with a certified reference value expressed in amount of DNA molecules per well. An interlaboratory study, conducted using blinded NA008 High GC reference material to assess reproducibility among seven independent laboratories demonstrated less than 4.5% reproducibility relative standard deviation. With the exclusion of one laboratory, laboratories had appropriate technical competency, fully functional instrumentation, and suitable reagents to perform accurate ddPCR based DNA quantification measurements at the time of the study. The study results confirmed that NA008 High GC reference material is fit for the purpose of being used for quality control of ddPCR systems, consumables, instrumentation, and workflow.

    Publication / Type:
    Analytical Chemistry
    Related Subject:
    DNA, ddPCR
    Link:
    http://pubs.acs.org/doi/10.1021/acs.analchem.6b05032
  • Synthesis, debugging, and effects of synthetic chromosome consolidation: synVI and beyond Leslie A. Mitchell, et. al.

    Institution: NYU School of Medicine, Boeke Lab

    Publication: Science

    2017 PDF abstract

    Total synthesis of designer chromosomes and genomes is a new paradigm for the study of genetics and biological systems. The Sc2.0 project is building a designer yeast genome from scratch to test and extend the lim- its of our biological knowledge. Here we describe the design, rapid assembly, and characterization of synthetic chromosome VI (synVI). Further, we investigate the phenotypic, transcriptomic, and some consolidation to uncover possible synthetic genetic interactions and/or perturbations of native cellular networks as the number of designer changes increases is the next major step for the Sc2.0 project.

    Publication / Type:
    Science
    Related Subject:
    Leslie A. Mitchell, Synthetic Biology, Genomics
    Link:
    http://science.sciencemag.org/content/355/6329/eaaf4831
  • Bug mapping and fitness testing of chemically synthesized chromosome X Yi Wu, et. al.

    Institution: NYU School of Medicine, Boeke Lab

    Publication: Science

    2017 PDF abstract

    Design and construction of an extensively modified yeast genome is a direct means to interrogate the integrity, com- prehensiveness, and accuracy of the knowl- edge amassed by the yeast community to date. The international synthetic yeast genome project (Sc2.0) aims to build an entirely de- signer, synthetic Saccharomyces cerevisiae ge- nome. The synthetic genome is designed to increase genome stability and genetic flexi- bility while maintaining cell fitness near that of the wild type. A major challenge for a ge- nome synthesis lies in identifying and elim- inating fitness-reducing sequence variants referred to as “bugs.”

    Publication / Type:
    Science
    Related Subject:
    Yi Wu, Synthetic Biology, Genomics
    Link:
    http://science.sciencemag.org/content/355/6329/eaaf4706
  • Precision Cancer Medicine in the Acoustic Dispensing Era: Ex Vivo Primary Cell Drug Sensitivity Testing Evgeny Kulesskiy, Jani Saarela, Laura Turunen, and Krister Wennerberg

    Institution: Institute for Molecular Medicine Finland (FIMM), University of Helsinki, Finland

    Publication: Journal of Laboratory Automation (JALA) Special Issue

    2016 PDF abstract

    Cancer therapy is increasingly becoming individualized, but there are also big gaps between the molecular knowledge of individual cancers we can generate today and what can be applied in the clinic. In an attempt to bridge this knowledge gap between cancer genetic and molecular profiling and clinically useful information, an individualized systems medicine program has been established at the Institute for Molecular Medicine Finland (FIMM), University of Helsinki, and the Helsinki University Hospital. Central to this program is drug sensitivity and resistance testing (DSRT), in which responses of primary cancer cells to a comprehensive clinical oncology and signal transduction drug collection are monitored. The drug sensitivity information is used with molecular profiling to establish hypotheses on individual cancer-selective targeting drug combinations and their predictive biomarkers, which can be explored in the clinic. Here, we describe how acoustic droplet ejection is enabling DSRT in our cancer individualized systems medicine program to (1) generate consistent but configurable assay-ready plates and determine how this affects data quality, (2) flexibly prepare drug combination testing plates, (3) dispense reagents and cells to the assay plates, and (4) perform ultra-miniaturized follow up assays on the cells from DSRT plates.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    automated biology, statistics, informatics and software, compound management, acoustic droplet ejection
    Link:
    http://jla.sagepub.com/content/21/1/27.full.pdf+html
  • Cell-Based High-Throughput Luciferase Reporter Gene Assays for Identifying and Profiling Chemical Modulators of Endoplasmic Reticulum Signaling Protein, IRE1 Juan Rong, et al.

    Institution: Cell Death and Survival Networks Program, Sanford-Burnham-Prebys Medical Discovery Institute

    Publication: J Biomol Screen

    2015 abstract

    Endoplasmic reticulum (ER) stress activates three distinct signal transducers on the ER membrane. Inositol-requiring protein 1 (IRE1), the most conserved signal transducer, plays a key role in ER stress-mediated signaling. During ER stress, IRE1 initiates two discrete signaling cascades: the "adaptive" signaling cascade mediated by the XBP1 pathway and the "alarm" signaling cascade mediated by stress-activated protein kinase pathways. Fine-tuning of the balance between the adaptive and alarm signals contributes significantly to cellular fate under ER stress. Thus, we propose that the design of high-throughput screening (HTS) assays to selectively monitor IRE1 mediated-signaling would be desirable for drug discovery. To this end, we report the generation of stable human neural cell lines and development of cell-based HTS luciferase (Luc) reporter gene assays for the identification of pathway-specific chemical modulators of IRE1. We implemented a cell-based Luc assay using a chimeric CHOP-Gal4 transcription factor in 384-well format for monitoring IRE1 kinase-mediated p38MAPK activation and an unfolded response pathway element (URPE)-Luc cell-based assay in 1536-well format for monitoring IRE1's RNase-mediated activation of XBP1. Chemical library screening was successfully conducted with both the CHOP/Gal4-Luc cells and UPRE-Luc engineered cells. The studies demonstrate the feasibility of using these HTS assays for discovery of pathway-selective modulators of IRE1.

    Publication / Type:
    J Biomol Screen
    Related Subject:
    ER stress, IRE1, XBP1, p38MAPK, high-throughput screen
    Link:
    http://jbx.sagepub.com/content/20/10/1232.abstract
  • Development of a Medium-Throughput Targeted LCMS Assay to Detect Endogenous Cellular Levels of Malonyl-CoA to Screen Fatty Acid Synthase Inhibitors Philip J. Hopcroft and David I. Fisher

    Institution: Discovery Sciences, AstraZeneca

    Publication: J Biomol Screen

    2015 abstract

    The fatty acid synthase (FAS) enzyme in mammalian cells is a large multidomain protein responsible for de novo synthesis of fatty acids. The steps catalyzed by FAS involve the condensation of acetyl-CoA and malonyl-CoA moieties in the presence of NADPH until palmitate is formed. Inhibition of FAS causes an accumulation of intracellular malonyl-CoA, as this metabolite is essentially committed to fatty acid synthesis once formed. Detection of intracellular metabolites for screening can be problematic due to a lack of appropriate tools, but here we describe a targeted liquid chromatography – mass spectroscopy (LCMS) method to directly measure endogenous levels of malonyl-CoA to drive a drug development structure–activity relationship (SAR) screening cascade. Our process involves preparation of samples at 96-well scale, normalization postpermeabilization via use of a whole-well imaging platform, and the LCMS detection methodology. The assay is amenable to multiplexing cellular endpoints, has a typical Z′ of >0.6, and has high reproducibility of EC50 values.

    Publication / Type:
    J Biomol Screen
    Related Subject:
    malonyl-CoA, fatty acid synthase, LCMS, screening
    Link:
    http://jbx.sagepub.com/content/early/2015/11/19/1087057115617894.abstract
  • Acoustic Methods to Monitor Protein Crystallization and to Detect Protein Crystals in Suspensions of Agarose and Lipidic Cubic Phase Daniel L. Ericson, Xingyu Yin, Alexander Scalia, Yasmin N. Samara, Richard Stearns

    Institution: Office of Educational Programs, Brookhaven National Laboratory

    Publication: Journal of Laboratory Automation (JALA) Special Issue

    2015 abstract

    Improvements needed for automated crystallography include crystal detection and crystal harvesting. A technique that uses acoustic droplet ejection to harvest crystals was previously reported. Here a method is described for using the same acoustic instrument to detect protein crystals and to monitor crystal growth. Acoustic pulses were used to monitor the progress of crystallization trials and to detect the presence and location of protein crystals. Crystals were detected, and crystallization was monitored in aqueous solutions and in lipidic cubic phase. Using a commercially available acoustic instrument, crystals measuring ~150 µm or larger were readily detected. Simple laboratory techniques were used to increase the sensitivity to 50 µm by suspending the crystals away from the plastic surface of the crystallization plate. This increased the sensitivity by separating the strong signal generated by the plate bottom that can mask the signal from small protein crystals. It is possible to further boost the acoustic reflection from small crystals by reducing the wavelength of the incident sound pulse, but our current instrumentation does not allow this option. In the future, commercially available sound emitting transducers with a characteristic frequency near 300 MHz should detect and monitor the growth of individual 3 µm crystals.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    crystal detection, lipidic cubic phase, crystallography, acoustic droplet ejection, crystallization, visualization, sonar
    Link:
    http://jla.sagepub.com/content/21/1/107.full.pdf+html
  • Maintaining Microclimates during Nanoliter Chemical Dispensations Using CustomDesigned Source Plate Lids B Foley, A Drozd, M Bollard, D Laspina, N Podobedov

    Institution: Office of Educational Programs, Brookhaven National Laboratory, Upton, NY

    Publication: Journal of Laboratory Automation (JALA) Special Issue

    2015 abstract

    A method is described for using custom snap-on lids to protect chemicals in microtiter plates from evaporation and contamination. The lids contain apertures (diameter 1.5, 1.0, or 0.5 mm) through which the chemical building blocks can be transferred. The lid with 0.5 mm apertures was tested using a noncontact acoustic liquid handler; the 1.0 and 1.5 mm lids were tested using two tip-based liquid handlers. All of the lids reduced the rate at which solvents evaporated to room air, and greatly reduced the rate of contamination by water and oxygen from room air. In steady-state measurements, the lids reduced the rate of evaporation of methanol, 1-hexene, and water by 33% to 248%. In cycled experiments, the contamination of aqueous solvent with oxygen was reduced below detectability and the rate at which DMSO engorged atmospheric water was reduced by 81%. Our results demonstrate that the lids preserve the integrity of air-sensitive reagents during the time needed for different types of liquid handlers to perform dispensations. Controlling degradation and evaporation of chemical building blocks exposed to the atmosphere is increasingly useful as the reagent volume is reduced by advances in liquid handling technology, such as acoustic droplet ejection.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    acoustic droplet ejection, liquid handling, chemistry, contamination, microdroplets, evaporation, dehydration, synthesis, solvents, crystallization, drug discovery
    Link:
    http://jla.sagepub.com/content/21/1/115.full.pdf+html
  • Moving Liquids with Sound: The Physics of Acoustic Droplet Ejection for Robust Laboratory Automation in Life Sciences Babur Hadimioglu, Richard Stearns, and Richard Ellson

    Institution: Labcyte Inc.

    Publication: Journal of Laboratory Automation (JALA) Special Issue

    2015 abstract

    Liquid handling instruments for life science applications based on droplet formation with focused acoustic energy or acoustic droplet ejection (ADE) were introduced commercially more than a decade ago. While the idea of “moving liquids with sound” was known in the 20th century, the development of precise methods for acoustic dispensing to aliquot life science materials in the laboratory began in earnest in the 21st century with the adaptation of the controlled “drop on demand” acoustic transfer of droplets from high-density microplates for high-throughput screening (HTS) applications. Robust ADE implementations for life science applications achieve excellent accuracy and precision by using acoustics first to sense the liquid characteristics relevant for its transfer, and then to actuate transfer of the liquid with customized application of sound energy to the given well and well fluid in the microplate. This article provides an overview of the physics behind ADE and its central role in both acoustical and rheological aspects of robust implementation of ADE in the life science laboratory and its broad range of ejectable materials.

    Publication / Type:
    Journal of Laboratory Automation (JALA) Special Issue
    Related Subject:
    acoustic droplet ejection, ADE, dynamic fluid analysis, DFA
    Link:
    http://jla.sagepub.com/content/21/1/4.full.pdf+html
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