Super-resolution Chromatin Visualization Using a Combined Method of Fluorescence In Situ Hybridization and Structured Illumination Microscopy in Solanum lycopersicum.
Super-resolution Chromatin Visualization Using a Combined Method of Fluorescence In Situ Hybridization and Structured Illumination Microscopy in Solanum lycopersicum.
复制标题
使用荧光原位杂交和结构照明显微镜相结合的方法对番茄进行超分辨率染色质可视化。
DOI：
10.1007/978-1-0716-2253-7_7
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Kuo P
中科院分区：
文献类型：
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作者：
Kuo P
染色质组织影响基因和转座子的表达，并调节各种细胞过程。高阶染色质结构已广泛研究使用基因组方法和显微镜图像分析。染色体构象捕获和DNA片段连接的测序使染色质相互作用和染色体折叠的研究成为可能。然而，某些细胞类型嵌入到其他细胞类型中，这使得使用高通量基因组方法研究它们的过程变得复杂。为了克服这一限制，高分辨率显微镜技术现在可用于研究单细胞中的染色质组织。在本章中，我们提供了一个详细的方案来准备染色体从番茄细胞核扩散，荧光原位杂交标记基因组位点，并在高分辨率下用结构照明显微镜观察这些位置。
Chromatin organization influences gene and transposon expression, and regulates various cellular processes. Higher order chromatin structure has been widely studied using genomic approaches and microscopy image analyses. Chromosome conformation capture and sequencing the junction of DNA fragments enables the study of both chromatin interaction and chromosome folding. However, certain cell types are embedded in other cell types which complicate the process of studying them using high-throughput genomic approaches. To overcome this limitation, high-resolution microscopy techniques are now available to investigate chromatin organization in single cells. In this chapter, we provide a detailed protocol to prepare chromosome spreading from tomato nuclei, to label genomic loci by fluorescence in situ hybridization, and to visualize these locations at high resolution with Structured Illumination microscopy.