Cell-based high content screening is becoming a widely used methodology in therapeutic drug discovery and functional genomics. Changes in cellular morphology and biomarker distributions in response to small molecules or gene knockdowns provide an information-rich profile of cellular responses. One limitation that exists with these cell-based assays is the availability of image processing algorithms that are capable of reliably and automatically analyzing HCS images. HCS images of primary neuron cell cultures are particularly challenging to analyze because of their complex morphology. My group developed a robust method for quantifying the morphology of neuronal cells in HCS images. The novelties of our method include its capability to deal with non-uniform illumination, detect faint neurites, and perform statistical analysis of morphological features. The power of our method was demonstrated in its applications to analyze neural outgrowth genes using genome-wide RNAi [1], screen chemical-compounds using primary neuron cell cultures derived from embryos of a Drosophila Huntington's Disease model [2], and study the detailed biological functions of a Drosophila protein in regulating synapse formation, axonal transport, and microtubule organization [3].

[1] Identification of neural outgrowth genes using genome-wide RNAi. Sepp KJ, Hong P, Lizarraga SB, Liu JS, Mejia LA, Walsh CA, Perrimon N. PLoS Genet, 2008. 4(7): p. e1000111.

[2] Automatic Robust Neurite Detection and Morphological Analysis of Optical Microscopy Neuron Cell Culture Images in High-Content Screening. Wu C, Schulte J, Sepp KJ, Littleton JT., and Hong P. Neuroinformatics (in press).

[3] DMob4/Phocein regulates synapse formation, axonal transport, and microtubule organization. Schulte J, Sepp KJ, Jorquera R, Wu C, Song Y, Hong P, and Littleton JT. Journal of Neuroscience (in press).

[4] Schoemans, R., M.S. Aigrot, C. Wu, R. Maree, P. Hong, S. Belachew, C. Josse, C. Lubetzki, and V. Bours, Oligodendrocyte development and myelinogenesis are not impaired by high concentrations of phenylalanine or its metabolites. J Inherit Metab Dis, 2010.

Examples

Original Images	Neurite Extraction Result