细菌细胞中小RNA的定量超分辨率成像

Quantitative Super-Resolution Imaging of Small RNAs in Bacterial Cells.

作者信息

Park Seongjin, Bujnowska Magda, McLean Eric L, Fei Jingyi

机构信息

Department of Biochemistry and Molecular Biology, The University of Chicago, Chicago, IL, USA.

The College of The University of Chicago, Chicago, IL, USA.

出版信息

Methods Mol Biol. 2018;1737:199-212. doi: 10.1007/978-1-4939-7634-8_12.

DOI:10.1007/978-1-4939-7634-8_12

PMID:29484595

Abstract

We present a method for the quantification of small regulatory RNAs (sRNAs) in bacteria, by combining single-molecule fluorescence in situ hybridization (smFISH), super-resolved single-fluorophore microscopy, and clustering analysis. Compared to smFISH imaging with diffraction-limited fluorescence microscopy, our method provides better quantification for short and abundant RNA (such as sRNAs) in a small volume of bacterial cells. Our method can also be directly used for the quantification of messenger RNAs (mRNAs).

摘要

我们提出了一种通过结合单分子荧光原位杂交（smFISH）、超分辨单荧光团显微镜和聚类分析来定量细菌中小调节RNA（sRNA）的方法。与使用衍射极限荧光显微镜的smFISH成像相比，我们的方法能更好地定量少量细菌细胞中的短且丰富的RNA（如sRNA）。我们的方法也可直接用于信使RNA（mRNA）的定量。