MetaXpress High-Content Image Acquisition and Analysis Software

In this application note, Molecular Devices describes a method for semi-automated culturing and monitoring of cerebral organoids, as well as the testing of functional neuronal activity by means of recording Ca2+ oscillations.

In this application note, Molecular Devices demonstrates how the ImageXpress® Micro Confocal system and MetaXpress® software allows rapid imaging and analysis of 3D spheroids in microplates for monitoring induced apoptosis and mitochondrial toxicity of anti-cancer drugs.

In this application note, Molecular Devices demonstrates the use of the Elplasia 96-well plates with a 3D culture workflow that includes spheroid generation, compound treatment, cytotoxicity assay, high-content imaging on the ImageXpress® Micro Confocal High-Content Imaging System, and 3D image analysis using MetaXpress® High-Content Image Acquisition and Analysis Software. The ability to easily generate multiple spheroids and to seamlessly integrate the workflow with high-content imaging promises to have significant applications in drug discovery and compound toxicology.

The focus of this application note is to develop a high-throughput 3D neurite outgrowth assay using iPSC-derived neurons developed in the microfluidic OrganoPlate® platform, with the goal of establishing 3D models for neurodegenerative diseases and neurotoxicology screens. The OrganoPlate® is a high-throughput platform that combines the most recent advances in 3D cell culture, Phaseguides™ and microfluidics. The OrganoPlate contains 96 tissue chips suitable for long-term culture of live cells, is amenable for screening purposes, and is compatible with standard laboratory equipment or automated systems, such as the ImageXpress® Micro Confocal High-Content Imaging System.

The new Curve Fitting Algorithms in the MetaXpress® 6.5 (MX 6.5) HighContent Image Acquisition and Analysis Software were designed to facilitate data visualization and the secondary analysis of data. The software allows you to generate a curve of best fit to a series of data points. With multiple functions, including 4-parameter logistic, and multiple curve outputs, such as EC50 and benchmark response, an array of graphs and curves can be generated to best represent data. Here, we show how to generate doseresponse relationship for defining the EC50 values of compounds using a toxicity assay as an example.

Cellular transformation/tumorigenicity assays using cultures of cells in semi-solid media (soft agar or Matrigel) has been well established for cancer research. The assay requires cells to grow in an anchorage independent manner, which is a hallmark of cancer cells. Compared with adherent cells grown in 2D monolayers, 3D growth conditions are believed to more accurately reflect the natural environment of cancer cells and span the gap between 2D cultures and animal studies. Importantly, 3D assays correlated better with tumorigenicity in animals, e.g. mouse xenografts. The growing interest in developing methods for personalized medicine, in which tumor cells from individual patients are tested for sensitivityto a panel of drugs , has led to a need for more relevant, time-sensitive studies.

There is increasing interest in exploring the use of three-dimensional (3D) spheroids for modeling developmental and tissue biology with the goal of accelerating translational research in these areas. As a result, the development of higher throughput quantitative assays using 3D cultures is an active area of investigation. In this study, we developed and optimized methods for the formation of 3D liver spheroids derived from human iPS cells as well as confocal imaging and analysis methods for toxicity assessment.

The ImageXpress® Micro Confocal/ XLS Widefield High-Content Imaging System from Molecular Devices includes optional Fluidics and Environmental Control modules, enabling single-channel pipetting for compound addition and media exchange during rapid kinetics or extended time-lapse experiments. The Fluidics option is powered by MetaXpress® High Content Image Acquisition and Analysis Software, allowing precise control of each pipetting event.

Screening of hybridoma supernatants for antibodies directed against cell-surface antigens is an important step in the discovery and development of antibodies for vaccines or therapeutic use. The sensitivity of these assays is increased by taking advantage of the optical properties of confocal imaging such as utilizing the narrow depth of field to acquire high signal-to-background, crisp images with no out of focus light creating image blur.

Label-free cellular assays are required for a multitude of biological applications that monitor the cell number, proliferation, health, confluency, and cytotoxicity. These applications necessitate efficient and robust transmitted light (TL) imaging and analysis capabilities providing precise segmentation for quantitation of cells and assessment in variety of cell responses and morphologies. Additionally, the coupling of high-contrast transmitted light imaging and fluorescent labeling and imaging is essential to numerous cellbased assays.