Dual-color dSTORM imaging and ThunderSTORM image reconstruction and analysis to study the spatial organization of the nuclear phosphatidylinositol phosphates.
Dual-color dSTORM imaging and ThunderSTORM image reconstruction and analysis to study the spatial organization of the nuclear phosphatidylinositol phosphates.
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DOI：
10.1016/j.mex.2021.101372
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发表时间：
2021
期刊：
影响因子：
1.9
通讯作者：
Hozák P
中科院分区：
文献类型：
--
作者：
Hoboth P;Šebesta O;Sztacho M;Castano E;Hozák P
实现并优化了核PIP2的双色dSTORM成像。利用ImageJ2插件雷暴自定义最近邻距离分析，定量评价核PIP2的空间分布。开发了ImageJ2工具，用于celllo中最近邻距离分析结果的可视化。单分子定位显微镜（SMLM）为蛋白质和核酸的亚核组织提供了前所未有的见解，但除了核包膜外，核脂质在细胞核功能组织中的作用研究较少。然而，核脂质，特别是磷脂酰肌醇磷酸（PIPs）在基因表达中发挥着越来越明显的作用。因此，在这里，我们提供了基于smlm的方法来定量评估核pip分布，同时保留核结构的背景。具体来说，以磷脂酰肌醇4,5-二磷酸（PIP2）为例，我们有：我们定制的双色dSTORM成像和定量分析提供了一种独立于核PIPs的生化和脂质组学分析但又互补的工具。与生化和脂质组学分析相反，我们分析的优势在于它保留了核PIP分布的空间背景。
Implemented and optimized the dual-color dSTORM imaging of nuclear PIP2. Customized the Nearest Neighbor Distance analysis using ImageJ2 plug-in ThunderSTORM to quantitatively evaluate the spatial distribution of nuclear PIP2. Developed an ImageJ2 tool for the visualization of the Nearest Neighbor Distance analysis results in cellulo. Single molecule localization microscopy (SMLM) provided an unprecedented insight into the sub-nuclear organization of proteins and nucleic acids but apart from the nuclear envelope the role of the nuclear lipids in the functional organization of the cell nucleus was less studied. Nevertheless, nuclear lipids and specifically phosphatidylinositol phosphates (PIPs) play increasingly evident roles in gene expression. Therefore, here we provide the SMLM-based approach for the quantitative evaluation of the nuclear PIPs distribution while preserving the context of nuclear architecture. Specifically, on the example of phosphatidylinositol 4,5-bisphosphate (PIP2) we have:Our customization of the dual-color dSTORM imaging and quantitative analysis provide a tool that is independent of but complementary to the biochemical and lipidomic analyses of the nuclear PIPs. Contrary to the biochemical and lipidomic analyses, the advantage of our analysis is that it preserves the spatial context of the nuclear PIP distribution.
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影响因子：
2.3
作者：
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DOI：
10.1016/j.bbalip.2021.158890
发表时间：
2021-02-13
影响因子：
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作者：
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通讯作者：
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