Accessible technology for high-content analysis
High-content analysis (HCA) integrates fluorescence microscopy, image processing, automated measurements, and informatics into a complex tool that has not only facilitated fundamental discoveries in biology but also enabled the progression of many compounds through the drug discovery process. Since their introduction in 1999, the Thermo Scientific™ ArrayScan™ HCA readers have been cited in over 1,000 publications for application areas such as oncology, neuroscience, and toxicology. HCA (also referred to as high-content screening or HCS) has become a staple of systems biology research, particularly when multiple cellular events are tracked in response to a stimulus or treatment.
Wanted: An advanced yet accessible HCA instrument
As researchers address a broader range of phenotypes with HCA, they need more capabilities from their hardware and software tools. Sample types were once limited to microscope slides and 96-well plates, but they now include 3D matrices and all SBS (Society of Biomolecular Screening) microwell formats. The biological content of the samples also demands a wider range of techniques, from confocal imaging to brightfield microscopy, each with multiple wavelength options and software tools for analysis.
In this changing application landscape, selecting an instrument platform is challenging for scientists who don’t want to limit the scope of their experiments. And because budgets are tight, many laboratories are reluctant to commit the resources necessary for top-of-the-line HCA platforms. The new Thermo Scientific™ Cellinsight™ CX7 High Content Analysis Platform is an integrated benchtop instrument that interrogates multiple sample types with a wide range of techniques, all within a physical and fiscal footprint to meet the needs of most laboratories (Figure 1).
Meet the Cellinsight CX7 HCA Platform
With the Cellinsight CX7 platform, your HCA assays can combine any of the imaging modes—brightfield, widefield, and confocal (Figure 2)—to extract the information you need from your samples. Each modality takes advantage of a proprietary laser-based autofocus technique that enables fast and reproducible read times, even when sample wells are sparsely populated.
For brightfield imaging, colorimetric absorbance measurements from histology samples are achieved using the LED array for RGB and amber illumination. Moreover, you can multiplex classic stains like hematoxylin and eosin (H&E) with fluorescent probes, offering new possibilities for data correlation in tissue sections. For confocal imaging, high-speed CrEST™ spinning-disk confocal technology with 40 μm or 70 μm pinholes is built into the optical path to provide sharp imaging in thick tissue samples and 3D matrices. The widefield imaging mode occupies the same light path as the confocal mode, sharing a 7-color solid-state LED light engine that provides a wide spectrum of excitation and maximizes your capabilities for performing multiplex assays. You can exploit the range of Molecular Probes™ fluorescent reagents—from HCS CellMask™ Blue stain to Alexa Fluor™ 750 dye—to minimize spectral overlap and improve assay quantitation.
A Photometrics™ X1 CCD camera with a 2,200 x 2,200 pixel array provides high sensitivity and resolution in all imaging modes. Together, the X1 camera and the 7-color light engine provide fast channel-switching times and minimize intensity fluctuations to help reduce scan times and boost quantitative performance. And getting started is easy: the Cellinsight CX7 platform is ready to accept screening plates from a robotic handler, or you can individually load your samples.
|Figure 2. Confocal image analysis of a fluorescently stained mouse kidney. A 16 μm cryostat section of mouse kidney was stained with Alexa Fluor™ 488 Wheat Germ Agglutinin (green), Alexa Fluor 568 Phalloidin (red), and DAPI (blue) and imaged on the Cellinsight CX7 HCA Platform using laser autofocus and confocal acquisition at 20x magnification: (A) nuclear identification and segmentation (blue); (B) phalloidin detection in WGA+ cells (red); (C) WGA+ cell selection (green); (D) WGA mask modification (green); (E) spot detection for quantifying WGA signal; (F) composite confocal image. (G) Scatter plot shows relative labeling intensity of the WGA vs. phalloidin conjugates in order to characterize phenotypes using intensity cutoffs.|
Efficiently analyze HCS data
For HCS to be truly accessible, it must meet the needs of novice users without sacrificing the power that experienced users have come to depend on. Thermo Scientific™ HCS Studio™ 3.0 Cell Analysis Software provides icon-driven guidance, starting with plate maps and annotation tools to set up assays and efficiently manage your experimental design. New users have a choice of more than 30 established assays that can be optimized to suit a particular cell line or phenotype. Using the intuitive icons, you can simply choose the assay and magnification of the optimized protocol, confirm settings, and begin to gather data on processes such as:
Experienced users can build their own assays from scratch using the flexible software tools. With instant feedback they can control hundreds of options, including:
For a researcher running assays on any scale, time-to-results is a critical measure. The sooner you have a result, the sooner you can make a decision, adjust a parameter, qualify an answer, or repeat an experiment. HCS Studio software users quickly recognize the benefits of intelligent software that processes data in real time, limiting acquisition to only the data required to generate a statistically significant result. For an assay like neurite outgrowth measurement in 96 wells, the Cellinsight CX7 platform can read the plate and report results in less than 4 minutes. Furthermore, the Assay Performance Tool of the HCS Studio software allows you to measure assay performance over multiple measurement criteria and select the parameters that give the best Z -prime result. At the end of the assay, you have results you can work with directly, without the delays of data spooling and off-line processing.
Explore your HCA options
The Cellinsight CX7 platform makes HCS accessible to more researchers by providing a range of high-performance tools in a convenient benchtop format. This technology platform builds on a 16-year legacy of HCS instrument and software development with ArrayScan readers and HCS Studio software, and almost 40 years of fluorescence expertise represented by Molecular Probes reagents.
For Research Use Only. Not for use in diagnostic procedures.