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Updated: Jun 10, 2026
08:18
Use of Single Molecule Fluorescent In Situ Hybridization SM-FISH to Quantify and Localize mRNAs in Murine Oocytes
Published on: April 24, 2019
10.8K
Whole-mount smFISH allows combining RNA and protein quantification at cellular and subcellular resolution
Lihua Zhao1, Alejandro Fonseca1, Anis Meschichi1
1Department of Plant Biology, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
Nature Plants
|June 15, 2023
Summary
Researchers developed a simple single-molecule RNA fluorescence in situ hybridization method for plants. This technique allows accurate mRNA counting and simultaneous protein detection in intact tissues at cellular resolution.
Area of Science:
- Plant biology
- Molecular biology
- Genetics
Background:
- Multicellular development relies on precise gene expression regulation.
- Quantifying mRNA in 3D plant tissues is difficult due to autofluorescence.
- Existing amplification-based methods are labor-intensive and prone to bias.
Purpose of the Study:
- To present a simplified method for absolute mRNA quantification in intact plant tissues.
- To enable simultaneous visualization of mRNA and protein levels at single-cell resolution.
- To overcome limitations of current in situ hybridization techniques in plants.
Main Methods:
- Single-molecule RNA fluorescence in situ hybridization (smFISH).
- Application to various intact plant tissues.
- Integration with fluorescent protein reporters for simultaneous mRNA and protein detection.
Main Results:
- Successful visualization and absolute counting of mRNA molecules in intact plant tissues.
- Simultaneous detection of mRNA and protein quantity and subcellular localization in single cells.
- Demonstration of a robust method for quantitative analysis of gene expression in plants.
Conclusions:
- This method simplifies the quantitative analysis of transcription and protein levels in plants.
- It provides cellular and subcellular resolution, crucial for understanding plant development.
- Enables researchers to fully leverage quantitative insights into gene and protein expression in plant tissues.
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