Category: High-Definition Biotechnology
High content analysis (HCA) is an automated imaging approach widely used for phenotypic screening in biological research and drug discovery, providing precise quantitation and spatial resolution of fluorescent or colorimetric signals from live and fixed biological specimens. Owing to needs for greater physiological relevance and modeling of living systems, the sample landscape has changed in recent years, shifting from traditional monolayer cultures to include increasingly complex preparations including 3D spheroids, patient derived stem cell lines and organoid cultures. While traditional HCA instrumentation implements widefield imaging with solid state LED or lamp based excitation for maximum throughput and analysis of traditional 2D cultures, such platforms lack the sensitivity and axial resolution needed to get meaningful data from 3D cultures. Here, we made a series of measurements from 2D and spheroid preparations, comparing confocal and widefield laser-based (LZR) excitation on the CellInsight CX7 HCA platform. Looking at measurements of cytoskeletal rearrangement, neurite outgrowth and cell survival in spheroid cultures, we found that confocal laser excitation gave improved axial resolution and penetrance of excitation light when making confocal measurements. Combining these capabilities with a functional on-stage incubator and intuitive software, we found these upgrades to the CX7 light engine greatly improved scan rates, spatial resolution and performance when collecting data from complex samples like 3D spheroids, primary neurons, and cytoskeletal rearrangement assays.
Daniel Beacham– Senior Staff Scientist, Thermo Fisher Scientific, Eugene, OR
Senior Staff Scientist
Thermo Fisher Scientific
Dan Beacham is a Senior Staff Scientist in the Discovery Biology group at the Molecular Probes Campus of Thermo Fisher Scientific in Eugene, Oregon. There, he develops cell-based imaging, high throughput screening and biochemical tools for drug discovery and basic sciences research. Dan comes to Thermo Fisher by way of Anchorage Alaska, with an Undergraduate Degree in Chemistry from Willamette University and a PhD in Neurobiology from Oregon Health and Sciences University, studying opioid regulation of voltage gated calcium ion channels under Edwin McCleskey. He came to Eugene in 2006 following Postdoctoral research at University College London and the Catterall laboratory in the Department of Pharmacology at the University of Washington in Seattle, where he studied molecular mechanisms of ion channel and GPCR control of neural excitability.