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Jan 1, 2011 (Vol. 31, No. 1)

Enabling Tools Extend the Range of HCA

Greater Heights Are Being Reached in Cell-Based Screening and Preclinical Analysis

  • Advancing High-Content Imaging

    Click Image To Enlarge +
    Field of view (FOV) comparison between a CCD and an sCMOS camera. [BioImaging Solutions]

    Dev Mittar, Ph.D., senior scientist at BD Biosciences, will present cell-based screening data for the identification and characterization of cell-surface markers for a monocyte-macrophage cell differentiation model using flow cytometry and high-content imaging. Together, says Dr. Mittar, “the two complementary technologies provide a more comprehensive single-cell analysis.”

    BD Pathway™ high-content cell analyzers are CCD camera-based automated cellular imaging systems that feature a selectable spinning disk that allows for confocal in addition to widefield imaging. The 435 instrument is a benchtop unit designed for performing fixed cell assays. The BD Pathway 855 system includes an environmental chamber with temperature and CO2 control to support live-cell imaging, including time-lapse studies, in 96- or 384-well plates. The 855 is capable of greater than four-color imaging. The stationary-stage design on both models allows cell samples to remain stationary while the objective moves across the plate. In this way, “you can image settled suspension cells without disturbing them,” says Dr. Mittar.

    BD developed the new Version 1.7 data-management tools for its AttoVision™ software to overcome the problems associated with data overload from high-content screening. The tools are part of a client/server-based system that allows multiple users to access data stored in a central repository, to add metadata to experimental results, and to select from a variety of query functions. The software extracts quantitative data from an image and then analyzes the data, generating graphs, charts, dose/response curves, and other forms of built-in or user-specified reporting tools.

    Dr. Mittar will also describe the company’s new BD Lyoplate™ Human Cell Surface Marker Screening Panel, which enables direct, antibody-based profiling of 242 cell-surface markers using either flow cytometry or high-content imaging.

  • Spinning-Disk Technology

    Click Image To Enlarge +
    High-content assay optimization for BD Lyoplate™ human cell surface marker screening: Representative pseudocolored merged images from antibody (green) and DAPI (blue) channels of differentiated THP-1 macrophages from control, CD11b (positive), and CD14 (negative) antibodies, respectively. Graph shows the average intensity quantified from antibody channels from control, CD11b, and CD14 wells (32 wells each) with Z'-factor values for the assay.

    Yokogawa Electric, through its western U.S. distributor BioImaging Solutions, will introduce its CellVoyager™ confocal high-content imaging systems, based on the company’s spinning-disk technology, to the U.S. and European markets in early 2011.

    Baggi Somasundaram, Ph.D., sales and marketing specialist for BioImaging Solutions, will demonstrate the capabilities of the benchtop CV1000 system, designed for the research market, during a technology showcase session at the meeting. He will also present the advantages of the CV6000 system, designed for high-throughput HCA in 6-, 24-, 96-, or 384-well plates for drug discovery screening applications.

    Dr. Somasundaram describes the CV1000 as a highly automated, benchtop confocal imaging system, with high precision and resolution that is ideal for basic research and assay development.

    The CV6000 offers the same high resolution as the CV1000 at higher throughput achieved through automation and multiple detectors. It can perform three-color imaging of a 96-well plate in one minute and a 384-well plate in five minutes. The CV600 images over a large coverage area “using an advanced five megapixel camera that captures an image field four times larger than the conventional CCD cameras currently used,” says Dr. Somasundaram.

    For live-cell imaging, both the CV1000 and CV6000 contain a built-in incubator chamber and feature automated X-Y stage adjustment to facilitate long-term observation of cells as they change over time and in response to the addition of a drug or introduction of other stimuli. The systems include the software needed to perform parallel data processing, and users can customize the data-analysis algorithms.

    Yokogawa recently announced a joint development agreement with the German Center for Neurodegenerative Diseases to collaborate on the development and application of cellular assays for HCA screening of compounds against neurodegenerative disease targets. Yokogawa will use this experience to enhance specific functions of the CV6000.

    Robert Graves, senior applications scientist at GE Healthcare Life Sciences (www.gelifesciences.com), will present the company’s new Zebrafish Analysis Plug-In for the IN Cell Investigator 1.6 software. The plug-in module enables automated organ-based analysis of zebrafish embryo images acquired from any microscope, and is optimized for use with GE’s In Cell Analyzer 2000 system, which performs whole-well imaging in 96-well plates.

    The system was designed for cellular assays and imaging of small organisms. Applications include testing of drug efficacy and toxicity. The In Cell Analyzer 2000 images a large field of view at high resolution, capturing an entire 96-well plate in a single image.

    Using transmitted light imaging, the system can produce label-free images of zebrafish embryos with sufficiently high resolution to enable organ recognition. Taking advantage of the well-defined and distinct shapes of individual zebrafish embryos, GE scientists created a flexible, geometric digital model that the software can use as a reference to identify specific organs.

    The built-in flexibility allows the software to make adjustments to the model to fit it to a particular embryo image; in this way, the software can make measurements even when the zebrafish is somewhat deformed. The IN Cell Analyzer 2000 can also obtain fluorescent images, and these can be linked to the digital model as well, thereby allowing quantification of fluorescence signals from different organs.

    What sets the Zebrafish Analysis Plug-in apart is “its organ-based approach,” says Ahmad Yekta, Ph.D., staff scientist at GE Healthcare Biosciences. It can identify 14 different organ regions, yielding 19 system-defined morphometric measurements including length and area. Users can also customize the software with additional measurements that will then automatically be collected, including measures of curvature, intensity, transparency, or granularity, for example.

    Mark Collins, global director of marketing for the Life Science Research-Cellomics business unit of Thermo Fisher Scientific (www.thermofisher.com), will introduce the company’s new personal cell-imaging platform, the CellInsight™, during the HCA technology showcase. Designed to overcome many of the barriers to entry into the field of high-content screening, the CellInsight incorporates many of the features of the company’s ArrayScan VTI HCS Reader and High Content Informatics (HCi) data-management and analysis platform.

    In particular, Collins points to the system’s ease of use, software-guided assay development feature, and solid-state construction with an optical/light train. The CellInsight has no moving parts except for the filter and stage. It is powered by a four-color LED engine, minimizing maintenance needs. For assay development, users select the appropriate algorithm and assay design modules built into the company’s iDev software, which interactively guides users through the process of training the algorithm to differentiate between positive and negative controls for a particular assay.

    Collins compares the cost of the system, about $100,000, to that of a high-end plate reader, making HCA more affordable and accessible for individual laboratories and researchers. Describing its speed he reports a “time to decision”—from image acquisition through data analysis—of about 3 minutes for a typical benchmark assay in a 96-well plate, less than 15 minutes for a 384-well plate, and about 60 minutes for a 1536-well plate.

    “When you want to scale a high-content assay and run it in different laboratories across an organization, a personal cell-imaging system solves the problem,” says Collins.


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