Abstract：
BACKGROUND:There is a need for effective computational methods for quantifying the three-dimensional (3-D) spatial distribution, cellular arbor morphologies, and the morphological diversity of brain astrocytes to support quantitative studies of astrocytes in health, injury, and disease. NEW METHOD:Confocal fluorescence microscopy of multiplex-labeled (GFAP, DAPI) brain tissue is used to perform imaging of astrocytes in their tissue context. The proposed computational method identifies the astrocyte cell nuclei, and reconstructs their arbors using a local priority based parallel (LPP) tracing algorithm. Quantitative arbor measurements are extracted using Scorcioni's L-measure, and profiled by unsupervised harmonic co-clustering to reveal the morphological diversity. RESULTS:The proposed method identifies astrocyte nuclei, generates 3-D reconstructions of their arbors, and extracts quantitative arbor measurements, enabling a morphological grouping of the cell population. COMPARISON WITH EXISTING METHODS:Our method enables comprehensive spatial and morphological profiling of astrocyte populations in brain tissue for the first time, and overcomes limitations of prior methods. Visual proofreading of the results indicate a >95% accuracy in identifying astrocyte nuclei. The arbor reconstructions exhibited 3.2% fewer erroneous jumps in tracing, and 17.7% fewer false segments compared to the widely used fast-marching method that resulted in 9% jumps and 20.8% false segments. CONCLUSIONS:The proposed method can be used for large-scale quantitative studies of brain astrocyte distribution and morphology.
journal_name
J Neurosci Methodsjournal_title
Journal of neuroscience methodsauthors
Kulkarni PM,Barton E,Savelonas M,Padmanabhan R,Lu Y,Trett K,Shain W,Leasure JL,Roysam Bdoi
10.1016/j.jneumeth.2015.02.014subject
Has Abstractpub_date
2015-05-15 00:00:00pages
38-51eissn
0165-0270issn
1872-678Xpii
S0165-0270(15)00066-7journal_volume
246pub_type
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journal_title：Journal of neuroscience methods
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