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Neurite outgrowth cell-by-cell analysis using the IN Cell Developer Toolbox

Key words: neurite outgrowth • cell-by-cell • fluorescent image analysis • IN Cell Analyzer • IN Cell Developer Toolbox


Neurite extension, or neurite outgrowth, plays a fundamental role in embryonic development, neuronal differentiation, and nervous system function. Neurite outgrowth is also critical in some neuropathological disorders as well as neuronal injury and regeneration. A variety of neurite parameters, including length and number of neurites, are sensitive to the extracellular environment and pharmacological agents. The cell-bycell analysis of neurite outgrowth with IN Cell Developer Toolbox allows quantitation of neurite extensions from fluorescently labeled cells imaged by an IN Cell Analyzer 1000 or 3000.

The protocol uses a two-color labeling approach, whereby the biological sample is labeled with two fluorophores that emit light at different wavelengths. One fluorophore is used to mark the cell nuclei and the other is used to identify the entire cytoplasm of each cell. The analysis procedure identifies nuclei, cell bodies, and neurites, discriminates between cellular components, and quantitates spatial characteristics of defined features.

Users can modify the protocol according to the requirements of a particular experiment: channels can be changed based on fluorescent markers used, parameters can be optimized according to cell size and fluorescence intensity, conditions for cell-by-cell segmentation of neurite networks can be defined as required, and parameters for data output can be chosen from the whole range of available measures.


Materials
Products used
IN Cell Analyzer 3000 25-8010-11

IN Cell Developer Toolbox Seat License 25-8098-26

Neurite Outgrow
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