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

Anna Maekiniemi, Robert H. Singer, Evelina Tutucci*

*Corresponding author for this work

Research output: Contribution to JournalArticleAcademicpeer-review

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
Article number105511
Pages (from-to)1-11
Number of pages11
JournalData in brief
Volume30
Early online date11 Apr 2020
DOIs
Publication statusPublished - Jun 2020

Funding

This work was supported by NIH Grant GM57071 to R.H.S. E.T was supported by Swiss National Science Foundation Fellowships P2GEP3_155692 and P300PA_164717 . We thank Ruth Hauptman for her help with the modification of the FISH-quant software.

FundersFunder number
National Institutes of HealthGM57071
Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen ForschungP300PA_164717, P2GEP3_155692

    Keywords

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

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