Single molecule mRNA fluorescent in situ hybridization combined with immunofluorescence in S. cerevisiae: Dataset and quantification

Anna Maekiniemi, Robert H. Singer, Evelina Tutucci

Research output: Contribution to journalArticle

1 Scopus citations

Abstract

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 S. cerevisiae. We measured the expression of the cell cycle-controlled mRNA CLN2 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 S. cerevisiae 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.

Original languageEnglish (US)
Article number105511
JournalData in Brief
Volume30
DOIs
StatePublished - Jun 2020

Keywords

  • Gene expression
  • Gene expression profiling
  • Imaging analysis
  • Immunofluorescence
  • Single cell
  • Single molecule mRNA FISH
  • smFISH
  • smFISH-IF

ASJC Scopus subject areas

  • General

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