Single molecule mRNA Fluorescent In Situ Hybridization combined to Immunofluorescence in S. cerevisiae: Dataset and quantification

Single molecule fluorescent in situ hybridization (smFISH) has emerged as a powerful technique that allows one to localize and quantify the absolute number of mRNAs in single cells. This approach has the potential to reveal the subcellular localization of mRNAs in individual cells, the cell-to-cell variability in gene expression and to precisely quantify the number of mRNAs in the cytoplasm and at the transcription site. In combination with immunofluorescence now smFISH can be used to correlate the expression on an mRNA of interest to the expression of a protein in single cells. The dataset provided here is described in the recently published smFISH-IF protocol “Simultaneous detection of mRNA and protein in S. cerevisiae by single molecule FISH and Immunofluorescence” [1]. We provide and quantify the smFISH-IF dataset detecting the cell cycle-controlled mRNA CLN2 and the cell cycle marker alpha-tubulin. In this article we analyse the smFISH data using the freely available Matlab-written software FISH-quant [2]. The provided datasets are meant to assist scientists interested in setting up smFISH-IF in their laboratory. Furthermore, software developers interested in creating imaging analysis tools for gene expression analysis in single cells, may find useful the provided the data.

单分子荧光原位杂交（single molecule fluorescent in situ hybridization，smFISH）已成为一种强大的实验技术，可实现对单个细胞内mRNA的定位及绝对数量的定量分析。该技术能够揭示单个细胞内mRNA的亚细胞定位、细胞间基因表达的异质性，并精准定量细胞质内以及转录位点处的mRNA数量。如今，将单分子荧光原位杂交与免疫荧光（immunofluorescence）技术联用，可在单个细胞中关联目标mRNA的表达与对应蛋白质的表达水平。
本数据集基于近期发表的单分子荧光原位杂交-免疫荧光（smFISH-IF）实验方案《通过单分子FISH与免疫荧光技术同步检测酿酒酵母（Saccharomyces cerevisiae，S. cerevisiae）中的mRNA与蛋白质》[1]。本数据集针对细胞周期调控的mRNA CLN2以及细胞周期标志物α微管蛋白（alpha-tubulin）进行了单分子荧光原位杂交-免疫荧光检测，并完成了定量分析。本文采用免费开源的MATLAB编写软件FISH-quant[2]对单分子荧光原位杂交数据进行分析。
本数据集旨在为有志于在实验室中搭建单分子荧光原位杂交-免疫荧光技术平台的科研人员提供协助。此外，对于致力于开发单细胞基因表达成像分析工具的软件开发人员而言，本数据集也具有较高的参考价值。