Cellomics HCS Reagent Kits for multiplex (4-parameter) analysis of cytotoxity.

Figure 1. Staining for nuclei, cell membrane permeability and mitochondrial transmembrane potential in HepG2 cells treated with media only or with 120 µM valinomycin for 24 hours. The cells were imaged using a Cellomics ArrayScan® HCS Reader.

Description

An essential aspect of all research is determining the toxic effects of compounds. Traditional toxicity screening methods use animal models which are expensive and have slow turnaround times. Furthermore, results obtained from animal models can be misleading because of differences between animals and humans.

The Multiparameter Cytotoxicity 2 HCS Reagent Kits contains optimized reagents for simultaneous detection and quantitation in the same cell of changes in nuclear morphology and size, the cell membrane’s permeability status, changes in cell density (number of cells per field) from compound toxicity, and changes induced to EITHER the lysosome’s mass or pH OR the mitochondria’s transmembrane potential. For these indicators of cell health, the supplied fluorescent reagents enable a fixed end-point assay. The assay has been developed, optimized and validated in the human hepatoma cell line HepG2 since any xenobiotic compound entering an animal is primarily targeted to the liver and kidney for metabolism and excretion. With minor protocol changes, this assay can be easily adapted for a variety of cell types or cell lines including those that originate from other xenobiotic targets like the lungs, central nervous system and heart. Once prepared cells can be analyzed using standard fluorescence microscopy or using a Cellomics ArrayScan HCS Reader with the Cell Health Profiling BioApplication, affording fully automated plate handling, focusing, image acquisition, analysis, quantification, and data storage.

This kit is available in three different versions. One version contains fluorescent probes for nuclear morphology/size (blue fluorescence), cell membrane’s permeability status (green fluorescence), lysosome mass or pH (orange fluorescence), and mitochondrial transmembrane potential (orange fluorescence). Because the probes for lysosomal mass and mitochondrial potential have similar spectra, this kit can be used to measure either organelle’s properties, depending on which fluorescent probe is used. The other two versions have, in addition to the nuclear and cell permeability probes, either the lysosomal or the mitochondrial fluorescent probe.

Features

• Available in multiplex or singleplex configurations
• Optimized protocol requiring no cell lysis, purification or filtration steps
• Immunofluorescence-based assay provides sensitive results without radioactivity
• Cells prepared with the HCS Reagent Kit may be analyzed directly via standard fluorescence microscopy; or HCS imaging systems
• Assay is versatile and widely applicable to a variety of cell types

Figure 2. Staining for nuclei, cell membrane permeability and lysosomal mass/pH in HepG2 cells treated with media only or with 100 µM tacrine for 24 hours. The cells were imaged using a Cellomics ArrayScan HCS Reader.

Related Poster Presentations

	Kay Opperman, Bhaskar S. Mandavilli, and Richik N. Ghosh (2008) Assessment of Oxidative Stress and DNA Damage in Individual Cells Using Automated, Quantitative Cell-Based Imaging.
	Bhaskar Mandavilli, Megan Cuda, Chandrasekaran Vasudevan, Richik N. Ghosh (2007) A New Panel of High Content Screening Cell-Based Assays for in vitro Cytotoxicity Measurements.
	Jeffrey R. Haskins, Bhaskar Mandavilli, Megan Cuda, Chandrasekaran Vasudevan, Richik N. Ghosh (2007) New reagent kits for high content cell-based assays for cytotoxicity.
	Chris Larson, Abe Dalu, Herve Lebrec, Jon Ingelfield (2006) Comparison of Manual Flask-Based vs. Cellomics Microplate-based Cytotoxicity Assay Methods.
	Chandrasekaran Vasudevan, Jeffrey A. Haskins, William E. Burmeister (2006) Assessing Toxicity of Nanoparticles with In Vitro Cell Based Assays.
	Vivek C. Abraham, Brent Samson, Oleg Lapets and Jeffrey R. Haskins (2004) Automated Correlative Analysis of Multiparametric Cellular Responses Measured using High Content Screening.
	Vivek C. Abraham, Brent Samson, Oleg Lapets and Jeffrey R. Haskins (2004) In Vitro Analysis of Multiparametric Cytotoxicity at the Individual Cell Level.
	Brent Samson, Vivek C. Abraham, Oleg Lapets and Jeffrey R. Haskins (2004) Use of Kinetic High Content Cell-based Assays for Rapid Profiling of Compound Toxicity.
	Richik N. Ghosh, Vivek Abraham, Linnette Grove, Oleg Lapets and Jeffrey R. Haskins (2003) A Kinetic High Content Screening Assay for Intracellular Compartmental Analysis in Single Cells.
	Vivek C. Abraham, Brent Samson, Oleg Lapets and Jeffrey R. Haskins (2003) Large-Scale Investigation of Cytotoxicity Enabled by Automated Simultaneous Monitoring of Multiple Intracellular Processes in Individual Living Cells.

Kit Contents

• Lysosomal mass/pH dye
• Mitochondrial membrane potential dye
• Hoechst dye permeability dye
• Wash buffer (10X Dulbecco's PBS)
• Thin plate seals

Ordering Information

Screening size kits and components (bulk and custom) available with special pricing. Please call.

To order Cellomics HCS Reagent Kits, please call 800-874-3723 in the U.S.
or contact your local distributor of Thermo Scientific Products.

Choose "Order Online" in table to see pricing and order online (U.S. only).

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