Cellomics HCS Reagent Kit for neuronal tubulin analysis of neuron cell bodies and neurites.

Description

Major efforts in CNS drug discovery research are focused on the identification of compounds that affect the growth of neurites. Drugs that promote nerve growth have potential curative effects in a wide variety of diseases and traumas that result in neuropathy and nerve injury, including stroke, spinal cord injuries, and neurodegenerative illnesses such as Parkinson’s and Alzheimer’s diseases. Current common methods for measuring neurite outgrowth include manual examination of individual cells under a microscope, and measurement of the total fluorescence from a labeled neuronal cell population using a fluorescence plate reader. The first of these methods is laborious and subjective; the second method cannon accurately discriminate between fluorescently labeled neuronal cell bodies and their neurites, inappropriately skewing data toward the brightly stained cell bodies.

The Neurite Outgrowth HCS Reagent Kit product provides a more effective solution for quantifying neurite outgrowth by combining high-content screening (HCS)-quality fluorescence reagents with a validated, automation-compatible protocol. Following stimulation of neurite outgrowth, cells growing in standard high-density microplates are fixed, permeabilized, and then fluorescently labeled. Nuclear staining (Hoechst Dye) identifies every cell within a population (Figure 1, left); an antibody specific to the neuronal form of tubulin labels neurons, neuronal-like cell bodies, and their neurites; the secondary antibody is conjugated with a fluorophore (Alexa Fluor® 488), resulting in the fluorescence of both the cell body and the neurites extending from it (Figure 1, right). The antibody is specific to neurons in a heterogeneous brain cell mixture, and can recognize neurons from a range of species. Thus, the time-consuming step of isolating neurons from a mixture of different cell types (i.e. primary brain cell cultures) is eliminated. Prepared cells can be analyzed using standard fluorescence microscopy or using a Cellomics fully automated HCS Reader with Neurite Outgrowth Bioapplication affording automated plate handling, focusing, image acquisition, analysis, quantification and data storage.

Features

• Neuronal-specific fluorescent reagent identifies neuronal cell bodies and neurites in a heterogeneous cell population without isolation steps
• Validated protocol for sample preparation & processing requires approximately 2.5h post-compound treatment and is automation compatible
• Cells prepared with the HCS Reagent Kit may be analyzed directly via standard fluorescence microscopy; or HCS imaging systems
• No radioactivity, cell lysis, purification, or filtration steps

Related Poster Presentations

	Amy M. Peters, Everett Ramer, Oleg Lapets, Lisa Reuter, Sarah Tencza, and Jeffrey Haskins (2006) Neuronal Morphology and Neurite Outgrowth Characterization Using An Automated Image Analysis System.
	Amy M. Peters, Everett Ramer, Oleg Lapets, Lisa Reuter, Sarah Tencza, and Jeffrey Haskins (2006) Neuronal Morphology and Neurite Outgrowth Characterization Using An Automated Image Analysis System.
	V. C. Abraham, R. N. Ghosh, P. Sammak, P. Petrosko and J. R. Haskins (2004) Application of High Content Screening to the Analysis of Stem Cell Growth & Differentiation.
	Everett Ramer, Stuart Arden, Richik Ghosh and Jeffrey R. Haskins (2003) Automated Cell-Based Quantitation of Neurite Outgrowth and Neuronal Cell Differentiation Using Cellomics Neurite Outgrowth BioApplications.

Kit Contents

• Neuronal-specific primary antibody
• Alexa Fluor® 488 conjugated secondary antibody
• Hoechst dye
• Wash buffer (10x)
• Permeabilization buffer (10x)
• Detergent buffer (10x)
• 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).

See all Cellomics HCS Reagent Kits

See product application note: A Live Cell, Kinetic, High Content Screening Application Interrogating Oxidative Stress in Neuroscreen-1™ Cells.