SAN FRANCISCO – Cambridge Healthtech Institute’s annual High-Content Analysis conference, held here this week, has traditionally served as an opportunity for players in the burgeoning high-content cell-based analysis and screening space to share data and present case studies in a relatively intimate atmosphere with highly focused presentations.
Many of these players — including tool vendors, pharmaceutical and biotech companies, and academic groups — present scientific posters detailing the use of high-content screening tools in drug discovery, functional genomics, and basic biomedical research.
A common theme of this year’s posters was how scientists are combining tools — including hardware, reagents, and software — from several different vendors, as opposed to a single high-content screening tool provider, to solve complex problems that demand a high degree of flexibility and customization.
This trend could be good news for tool vendors wanting a piece of the action. Although most of the original independent HCS instrument platform vendors have been assimilated into larger companies over the past five years, more and more small shops are opening up to offer specialized reagents, image-analysis software, and data-management tools for HCS.
Following is a recap of several notable poster presentations consistent with this theme.
TITLE: “High-content imaging-based study of the kinetics of cytoskeletal reorganization.”
PRESENTER: Aerie Pharmaceuticals
LEAD AUTHOR: Carmen Laethem
SUMMARY: Researchers from Aerie Pharmaceuticals studied the kinetics of cytoskeletal reorganization in porcine trabecular meshwork cells isolated from porcine eyes. Specifically, they examined changes in actin fiber length and the total area of focal adhesions using fluorescence reagents from Invitrogen, and the GE IN Cell 1000 platform running custom algorithms developed with the IN Cell Developer Toolbox v1.6.
TITLE: “Employing multi-parametric high-content cellular analysis to enrich lead identification and optimization at AstraZeneca.”
LEAD AUTHOR: Vicki Racicot
SUMMARY: AstraZeneca provided an overview of the variety of high-content screening assays the company is implementing globally, underscoring the fact that demand for specific HCS tools varies from lab to lab even within the same organization, depending on the applications being pursued. Specifically, AstraZeneca is using the Cellomics ArrayScan and Molecular Devices ImageXpress in Boston; the ArrayScan in Wilmington, Del.; the ArrayScan, GE Healthcare IN Cell Analyzer, and ImageXpress in Charnwood, UK; the ArrayScan, TTP Labtech Acumen Explorer, and IN Cell Analyzer in Alderley Park, UK; the ImageXpress in Sodertalje, Sweden; the ArrayScan in Lund, Sweden, and Molndal, Sweden; and the Evotec Technologies (now PerkinElmer) Opera in Brisbane, Australia.
TITLE: “High-throughput analysis of ligand, mutant, and kinase-dependent factors that affect androgen receptor function.”
PRESENTER: BaylorCollege of Medicine
LEAD AUTHOR: Adam Szafran
SUMMARY: Researchers from Michael Mancini’s group at Baylor College of Medicine have developed a single-cell assay that allows for rapid characterization of ligand-dependent cellular distribution of wild-type and mutant androgen receptor, and they relate this directly to transcriptional activity. To do this, they used the IC 100 Imaging Cytometer (formerly marketed by Beckman Coulter and currently serviced by Cellomics) and the Broad Institute’s CellProfiler image-analysis software.
TITLE: “Internalization of the CCR5 receptor by the antagonist met-RANTES – a role for endogenous aminopeptidases.”
PRESENTER: The Eskitis Institute for Cell and Molecular Therapies at Griffith University, Australia
LEAD AUTHOR: James Longden
SUMMARY: Developed a high-throughput screen for the effects of the CCR5 antagonist met-RANTES on receptor internalization in CHO cells with GFP-tagged CCR5 receptors. To do this, the researchers used the Evotec Technologies (now PerkinElmer) Opera high-content imager and Acapella imaging software.
TITLE: “A genome-wide siRNA screen for enhancers and suppressors of kinesin-5 activity during mitosis.”
PRESENTER:HarvardMedicalSchool department of systems biology
LEAD AUTHOR: Melody Tsui
SUMMARY: Researchers in the lab of Tim Mitchison tracked cellular responses to the combination of siRNA and drug addition using live-cell imaging on Molecular Devices’ ImageXpress MICRO imaging platform, with the Broad Institute’s CellProfiler and CellVisualizer open-source free software. They expect to use the screen to identify novel proteins required for cancer cell division and death.
TITLE: “Single cell-based multiplexed assay for phenotypic HCS for kinase inhibitors.”
PRESENTER: Merck KgaA
LEAD AUTHOR: Natalia Novac
SUMMARY: The researchers demonstrated a rapid method for determining pathway specificity of kinase inhibitors in secondary screening implementing anti-phospho-specific antibodies and fluorescently labeled secondary antibodies in multiplexed single cell-based analysis. They used the TTP Labtech Acumen Explorer microplate cytometer.
TITLE: “Strategies for promoting regeneration after CNS injury.”
PRESENTER: University of Miami, MillerSchool of Medicine
LEAD AUTHOR: John Bixby
SUMMARY: The researchers developed a phenotypic assay to examine whether any differentially expressed genes (i.e., cDNA library overexpression) could overcome the inhibitory effects of myelin in the hopes of identifying genes associated with the regeneration of neurons. They used the Cellomics KineticScan and Spotfire DecisionSite data-analysis software.
LEAD AUTHOR: Gareth Phillips
SUMMARY: Researchers used a Cellomics KineticScan to conduct a 96-well assay to detect three known toxic effects of nucleoside analogues on living cells. Nucleoside analogues are a major class of antivirals used to treat HIV, and are being explored for the treatment of hepatitis B and C.