An optimized fixation method containing glyoxal and paraformaldehyde for imaging nuclear bodies

doi:10.1261/rna.078671.120

	An optimized fixation method containing glyoxal and paraformaldehyde for imaging nuclear bodies
	Yao, Run-Wen 1; Luan, Peng-Fei 1; Chen, Ling-Ling1,2,3
	2021-06
发表期刊	RNA (IF:4.2[JCR-2023],4.4[5-Year])
ISSN	1355-8382
EISSN	1469-9001
卷号	27 期号:6 页码:725-733
DOI	10.1261/rna.078671.120
摘要	The mammalian cell nucleus contains different types of membrane-less nuclear bodies (NBs) consisting of proteins and RNAs. Microscopic imaging has been widely applied to study the organization and structure of NBs. However, current fixation methods are not optimized for such imaging: When a fixation method is chosen to maximize the quality of the RNA fluorescence in situ hybridization (FISH), it often limits the labeling efficiency of proteins or affects the ultrastructure of NBs. Here, we report that addition of glyoxal (GO) into the classical paraformaldehyde (PFA) fixation step not only improves FISH signals for RNAs in NBs via augmented permeability of the fixed nucleus and enhanced accessibility of probes, but also largely preserves protein fluorescent signals during fixation and immunostaining. We also show that GO/PFA fixation enables the covisualization of different types of nuclear bodies with minimal impact on their ultrastructures under super-resolution microscopy.
关键词	glyoxal paraformaldehyde nuclear body RNA fluorescence in situ hybridization immunofluorescence
收录类别	SCIE
语种	英语
WOS研究方向	Biochemistry & Molecular Biology
WOS类目	Biochemistry & Molecular Biology
WOS记录号	WOS:000652823600008
出版者	COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
原始文献类型	Article
引用统计	正在获取...
文献类型	期刊论文
条目标识符	https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/126792
专题	生命科学与技术学院_特聘教授组_陈玲玲组
通讯作者	Chen, Ling-Ling
作者单位	1.Chinese Acad Sci, Univ Chinese Acad Sci, Shanghai Inst Biochem & Cell Biol,State Key Lab M, CAS Ctr Excellence Mol Cell Sci,Shanghai Key Lab, Shanghai 200031, Peoples R China; 2.ShanghaiTech Univ, Sch Life Sci & Technol, Shanghai 201210, Peoples R China; 3.Univ Chinese Acad Sci, Hangzhou Inst Adv Study, Sch Life Sci, Hangzhou 330106, Peoples R China
通讯作者单位	生命科学与技术学院
推荐引用方式 GB/T 7714	Yao, Run-Wen,Luan, Peng-Fei,Chen, Ling-Ling. An optimized fixation method containing glyoxal and paraformaldehyde for imaging nuclear bodies[J]. RNA,2021,27(6):725-733.
APA	Yao, Run-Wen,Luan, Peng-Fei,&Chen, Ling-Ling.(2021).An optimized fixation method containing glyoxal and paraformaldehyde for imaging nuclear bodies.RNA,27(6),725-733.
MLA	Yao, Run-Wen,et al."An optimized fixation method containing glyoxal and paraformaldehyde for imaging nuclear bodies".RNA 27.6(2021):725-733.