ArticleJournal of visualized experiments : JoVE2013
Visualization and analysis of mRNA molecules using fluorescence in situ hybridization in Saccharomyces cerevisiae.
Article in Journal of visualized experiments : JoVE, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed.
- Modern RNA Quantification Methods: From RT-qPCR to Advanced Microscopy.The journal of physical chemistry. B · 2026Review
- Transcription factor localization dynamics and DNA binding drive distinct promoter interpretations.Cell reports · 2023Article
- European Society of Toxicologic Pathology (Pathology 2.0 Molecular Pathology Special Interest Group): Review of In Situ Hybridization Techniques for Drug Research and Development.Toxicologic pathology · 2023Review
- Strain-dependent differences in coordination of yeast signalling networks.The FEBS journal · 2023Article
- Pulsatile inputs achieve tunable attenuation of gene expression variability and graded multi-gene regulation.Nature communications · 2018Article
- An Optogenetic Platform for Real-Time, Single-Cell Interrogation of Stochastic Transcriptional Regulation.Molecular cell · 2018Article
- Systematic identification of factors mediating accelerated mRNA degradation in response to changes in environmental nitrogen.PLoS genetics · 2018Article
- Single-cell RNA sequencing reveals intrinsic and extrinsic regulatory heterogeneity in yeast responding to stress.PLoS biology · 2017Article
- Translation initiation events on structured eukaryotic mRNAs generate gene expression noise.Nucleic acids research · 2017Article
- Article
- The lncRNA RZE1 Controls Cryptococcal Morphological Transition.PLoS genetics · 2015Article
- Quantitative measurement of cancer tissue biomarkers in the lab and in the clinic.Laboratory investigation; a journal of technical methods and pathology · 2015Review
- Aro: a machine learning approach to identifying single molecules and estimating classification error in fluorescence microscopy images.BMC bioinformatics · 2015Article
- Genetic basis for Saccharomyces cerevisiae biofilm in liquid medium.G3 (Bethesda, Md.) · 2014Article
- Rapid synthesis and screening of chemically activated transcription factors with GFP-based reporters.Journal of visualized experiments : JoVE · 2013Article
Corrections and comments
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Authors and funding
7 authors.
Funding
Abstract
The Fluorescence in situ Hybridization (FISH) method allows one to detect nucleic acids in the native cellular environment. Here we provide a protocol for using FISH to quantify the number of mRNAs in single yeast cells. Cells can be grown in any condition of interest and then fixed and made permeable. Subsequently, multiple single-stranded deoxyoligonucleotides conjugated to fluorescent dyes are used to label and visualize mRNAs. Diffraction-limited fluorescence from single mRNA molecules is quantified using a spot-detection algorithm to identify and count the number of mRNAs per cell. While the more standard quantification methods of northern blots, RT-PCR and gene expression microarrays provide information on average mRNAs in the bulk population, FISH facilitates both the counting and localization of these mRNAs in single cells at single-molecule resolution.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.