Collar occupancy: A new quantitative imaging tool for morphometric analysis of oligodendrocytes
Collar occupancy: A new quantitative imaging tool for morphometric analysis of oligodendrocytes
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DOI：
10.1016/j.jneumeth.2017.11.014
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发表时间：
2018-01-15
影响因子：
3
通讯作者：
Domingues, Helena Sofia
中科院分区：
文献类型：
--
作者：
Boucanova, Filipa;Maia, Andre Filipe;Domingues, Helena Sofia
背景：少突胶质细胞是中枢神经系统的髓鞘细胞。从少突胶质前体细胞(OPC)分化为OL伴随着典型的形态变化。在OPC分化过程中，这些形态变化的定量成像通常用于表征细胞分化和髓鞘形成的新分子，以及筛选新的亲髓鞘药物。新方法：本文描述了一种自动化的高通量定量图像分析方法--衣领占有率，该方法允许以高含量的分析格式对体外培养的OL分化的不同阶段进行形态计量排名。衣领占有率是基于OPC/OL细胞质突起在包含突起网络的特定区域内所占面积的百分比来确定的。结果：我们观察到分化越多的细胞有更高的衣领占有率，因此，这一参数与OL的分化程度有关。与现有方法的比较：与人工分类方法相比，我们发现衣领占有率更稳健和无偏倚。此外，结合髓鞘碱性蛋白(MBP)染色定量髓鞘细胞的百分率，我们能够评估新的分子在OL分化和髓鞘形成中的作用，如Dusp19和Kank2。结论：我们成功地建立了一种自动化和定量的方法来表征OL体外分化的形态特征，可用于OL生物学的多种研究。(C)2017爱思唯尔B.V.保留所有权利。
Background: Oligodendrocytes (OL) are the myelinating cells of the central nervous system. OL differentiation from oligodendrocyte progenitor cells (OPC) is accompanied by characteristic stereotypical morphological changes. Quantitative imaging of those morphological alterations during OPC differentiation is commonly used for characterization of new molecules in cell differentiation and myelination and screening of new pro-myelinating drugs. Current available imaging analysis methods imply a non automated morphology assessment, which is time-consuming and prone to user subjective evaluation.New method: Here, we describe an automated high-throughput quantitative image analysis method entitled collar occupancy that allows morphometric ranking of different stages of in vitro OL differentiation in a high-content analysis format. Collar occupancy is based on the determination of the percentage of area occupied by OPC/OL cytoplasmic protrusions within a defined region that contains the protrusion network, the collar.Results: We observed that more differentiated cells have higher collar occupancy and, therefore, this parameter correlates with the degree of OL differentiation. Comparison with existing methods: In comparison with the method of manual categorization, we found the collar occupancy to be more robust and unbiased. Moreover, when coupled with myelin basic protein (MBP) staining to quantify the percentage of myelinating cells, we were able to evaluate the role of new molecules in OL differentiation and myelination, such as Dusp19 and Kank2.Conclusions: Altogether, we have successfully developed an automated and quantitative method to morphologically characterize OL differentiation in vitro that can be used in multiple studies of OL biology. (C) 2017 Elsevier B.V. All rights reserved.