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