Automated image analysis methods for 3-D quantification of the neurovascular unit from multichannel confocal microscope images
Automated image analysis methods for 3-D quantification of the neurovascular unit from multichannel confocal microscope images
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DOI：
10.1002/cyto.a.20149
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发表时间：
2005-07-01
期刊：
影响因子：
3.7
通讯作者：
Roysam, B
中科院分区：
文献类型：
--
作者：
Lin, G;Bjornsson, CS;Roysam, B
背景：需要综合和定量的方法来研究复杂系统元素之间的结构和功能关系，如神经血管单元（NVU），涉及多种细胞类型、微血管和各种基因组/蛋白质组学/离子功能实体。方法：血管铸型和选择性标记可以同时对微血管、细胞核和细胞质染色进行三维成像。多维分割是通过(1)滤出去除和衰减校正实现的；（ii）每个校正通道的独立分割和形态测量；(3)跨通道的空间关联特征计算。结合测量使细胞分类基于核形态测量，细胞质信号，和血管元件的距离。NVU元素之间的具体空间关系可以量化。结果：构建并验证了核与血管分割码与关联特征相结合的软件系统。生物变异导致了细胞核的错误识别（9.3%），细胞核的不分段（3.7%），细胞核的过分段（14%）和缺失（4.7%）。微血管分割错误很少发生，主要是由于管腔染色不均匀。结论：通过荧光通道的结合特征，结合标准特征，可以对NVU进行综合结构和功能分析。通过标记额外的结构和功能实体，该方法可以扩展到更大规模的系统生物学研究，整合空间和分子信息。(c) 2005 Wiley-Liss, Inc。
Background: There is a need for integrative and quantitative methods to investigate the structural and functional relations among elements of complex systems, such as the neurovascular unit (NVU), that involve multiple cell types, microvasculatures, and various genomic/proteomic/ionic functional entities.Methods: Vascular casting and selective labeling enabled simultaneous three-dimensional imaging of the microvasculature, cell nuclei, and cytoplasmic stains. Multidimensional segmentation was achieved by (i) bleed-through removal and attenuation correction; (ii) independent segmentation and morphometry for each corrected channel; and (iii) spatially associative feature computation across channels. The combined measurements enabled cell classification based on nuclear morphometry, cytoplasmic signals, and distance from vascular elements. Specific spatial relations among the NVU elements could be quantified.Results: A software system combining nuclear and vessel segmentation codes and associative features was constructed and validated. Biological variability contributed to misidentified nuclei (9.3%), undersegmentation of nuclei (3.7%), hypersegmentation of nuclei (14%), and missed nuclei (4.7%). Microvessel segmentation errors occurred rarely, mainly due to nonuniform lumen staining.Conclusions: Associative features across fluorescence channels, in combination with standard features, enable integrative structural and functional analysis of the NVU. By labeling additional structural and functional entities, this method can be scaled up to larger-scale systems biology studies that integrate spatial and molecular information. (c) 2005 Wiley-Liss, Inc.