Automated analysis of neurite branching in cultured cortical neurons using HCA‐Vision

Manual neuron tracing is a very labor‐intensive task. In the drug screening context, the sheer number of images to process means that this approach is unrealistic. Moreover, the lack of reproducibility, objectivity, and auditing capability of manual tracing is limiting even in the context of smaller studies. We have developed fast, sensitive, and reliable algorithms for the purpose of detecting and analyzing neurites in cell cultures, and we have integrated them in software called HCA‐Vision, suitable for the research environment. We validate the software on images of cortical neurons by comparing results obtained using HCA‐Vision with those obtained using an established semi‐automated tracing solution (NeuronJ). The effect of the Sez‐6 deletion was characterized in detail. Sez‐6 null neurons exhibited a significant increase in neurite branching, although the neurite field area was unchanged due to a reduction in mean branch length. HCA‐Vision delivered considerable speed benefits and reliable traces. © 2007 International Society for Analytical Cytology.

[1]  Kristel Michielsen,et al.  Morphological image analysis , 2000 .

[2]  J. Pierri,et al.  Altered cortical glutamate neurotransmission in schizophrenia: evidence from morphological studies of pyramidal neurons. , 2003, Annals of the New York Academy of Sciences.

[3]  W. Raub.  From the National Institutes of Health. , 1990, JAMA.

[4]  Paul J. Harrison.  The neuropathology of schizophrenia , 2008 .

[5]  Lily Yeh Jan,et al.  The Control of Dendrite Development , 2003, Neuron.

[6]  Peter Davies,et al.  Dendritic changes in Alzheimer's disease and factors that may underlie these changes , 1998, Progress in Neurobiology.

[7]  Changming Sun,et al.  Fast Linear Feature Detection Using Multiple Directional Non-Maximum Suppression , 2009, 18th International Conference on Pattern Recognition (ICPR'06).

[8]  Badrinath Roysam,et al.  Automated semantic analysis of changes in image sequences of neurons in culture , 2006, IEEE Transactions on Biomedical Engineering.

[9]  Joanne M. Britto,et al.  Sez-6 Proteins Affect Dendritic Arborization Patterns and Excitability of Cortical Pyramidal Neurons , 2007, Neuron.

[10]  J. Bilsland,et al.  A rapid method for semi-quantitative analysis of neurite outgrowth from chick DRG explants using image analysis , 1999, Journal of Neuroscience Methods.

[11]  Xiaobo Zhou,et al.  Automated neurite labeling and analysis in fluorescence microscopy images , 2006, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[12]  Changming Sun,et al.  Circular shortest path in images , 2003, Pattern Recognit..

[13]  B. Kampa,et al.  Synaptic integration in dendritic trees. , 2005, Journal of neurobiology.

[14]  F. Murakami,et al.  Growth cones of dorsal root ganglion but not retina collapse and avoid oligodendrocytes in culture. , 1995, Developmental biology.

[15]  E Meijering,et al.  Design and validation of a tool for neurite tracing and analysis in fluorescence microscopy images , 2004, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[16]  H. Moser,et al.  Dendritic anomalies in disorders associated with mental retardation. , 1999, Cerebral cortex.

[17]  Guo-li Ming,et al.  New directions in neuroregeneration , 2006, Expert opinion on biological therapy.

[18]  R. Wong,et al.  Activity-dependent regulation of dendritic growth and patterning , 2002, Nature Reviews Neuroscience.

[19]  J. Pierri,et al.  Altered Cortical Glutamate Neurotransmission in Schizophrenia , 2003 .