Quantitative 3-D analysis of GFAP labeled astrocytes from fluorescence confocal images
Quantitative 3-D analysis of GFAP labeled astrocytes from fluorescence confocal images
复制标题
DOI：
10.1016/j.jneumeth.2015.02.014
复制
发表时间：
2015-05-15
影响因子：
3
通讯作者：
Roysam, Badrinath
中科院分区：
文献类型：
--
作者：
Kulkarni, Prathamesh M.;Barton, Emily;Roysam, Badrinath
背景资料：有必要为定量的三维（3-D）空间分布，细胞乔木形态，和脑星形胶质细胞的形态多样性，以支持定量研究星形胶质细胞在健康，损伤和diseases.New方法的有效的计算方法：多重标记（GFAP，DAPI）的脑组织的共聚焦荧光显微镜被用来执行在其组织背景下的星形胶质细胞的成像。所提出的计算方法确定星形胶质细胞的细胞核，并重建其乔木使用本地优先级为基础的并行（LPP）跟踪算法。定量乔木测量提取使用Scorcioni的L-措施，并配置通过无监督谐波co-clustering揭示形态diversity.Results：所提出的方法确定星形胶质细胞核，产生3-D重建他们的乔木，并提取定量乔木测量，使细胞群体的形态分组。我们的方法首次实现了脑组织中星形胶质细胞群体的全面空间和形态学分析，并克服了现有方法的局限性。结果的视觉校对表明鉴定星形胶质细胞核的准确率>95%。乔木重建表现出减少3.2%的错误跳跃跟踪，和17.7%的假段相比，广泛使用的快速行进的方法，导致9%的跳跃和20.8%的假segments.Conclusions：所提出的方法可用于大规模的定量研究脑星形胶质细胞的分布和形态。(C)2015 Elsevier B. V.版权所有。
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. (C) 2015 Elsevier B.V. All rights reserved.