Maekiniemi Anna, Singer Robert H, Tutucci Evelina
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY 10461, United States.
Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY 10461, United States.
Data Brief. 2020 Apr 11;30:105511. doi: 10.1016/j.dib.2020.105511. eCollection 2020 Jun.
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. In combination with immunofluorescence (IF), smFISH can be used to correlate the expression of an mRNA and a protein of interest in single cells. Here, we provide and quantify an smFISH-IF dataset in We measured the expression of the cell cycle-controlled mRNA and the cell cycle marker alpha-tubulin. The smFISH-IF protocol describing the dataset generation is published in the accompanying article "Simultaneous detection of mRNA and protein in by single-molecule FISH and Immunofluorescence" [1]. Here, we analyze the smFISH data using the freely available software FISH-quant [2]. The provided datasets are intended to assist scientists interested in setting up smFISH-IF protocol in their laboratory. Furthermore, scientists interested in the generation of imaging analysis tools for single-cell approaches may find the provided dataset useful. To this end, we provide the differential interference contrast (DIC) channel, as well as multicolor, raw Z-stacks for smFISH, IF and DAPI.
单分子荧光原位杂交(smFISH)已成为一种强大的技术,可用于在单细胞中定位和定量mRNA的绝对数量。结合免疫荧光(IF),smFISH可用于关联单细胞中感兴趣的mRNA和蛋白质的表达。在此,我们提供并量化了一个smFISH-IF数据集。我们测量了细胞周期调控的mRNA以及细胞周期标记物α-微管蛋白的表达。描述数据集生成的smFISH-IF方案发表在随附文章《通过单分子FISH和免疫荧光同时检测mRNA和蛋白质》[1]中。在此,我们使用免费软件FISH-quant [2]分析smFISH数据。提供的数据集旨在帮助有兴趣在其实验室中建立smFISH-IF方案的科学家。此外,对单细胞方法的成像分析工具生成感兴趣的科学家可能会发现提供的数据集有用。为此,我们提供了微分干涉对比(DIC)通道以及用于smFISH、IF和DAPI的多色原始Z-stack。