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Updated: May 19, 2026
09:06
MicroRNA Amplification and Recognition through Locked-nucleic-acid In situ Hybridization as a Novel Detection and Quantification Method
Published on: October 7, 2025
Detection of MicroRNAs in Skin by In Situ Hybridization
Maximilian E Pickup1, Mohammed I Ahmed2
1Nottingham Trent University, School of Science and Technology, Nottingham, UK.
Methods in Molecular Biology (Clifton, N.J.)
|May 18, 2026
Summary
This study presents a detailed in situ hybridization (ISH) protocol to detect microRNAs (miRNAs) in skin and hair follicles. The method enables precise analysis of miRNA spatiotemporal expression patterns, overcoming previous technical challenges.
Area of Science:
- Molecular Biology
- Genetics
- Biochemistry
Background:
- MicroRNAs (miRNAs) are small noncoding RNAs regulating gene expression in animals and plants.
- Over one-third of protein-encoding mRNAs are estimated to be miRNA targets.
- Accurate determination of miRNA spatiotemporal expression is crucial but technically challenging.
Purpose of the Study:
- To establish and detail a robust in situ hybridization (ISH) protocol for miRNA detection.
- To enable the analysis of miRNA spatiotemporal expression patterns in skin and hair follicles.
- To provide a reliable method for miRNA localization in both frozen and paraffin-embedded tissues.
Main Methods:
- Detailed description of an in situ hybridization (ISH) protocol using LNA double-labeled probes.
- Application of digoxigenin-labeled probes for miRNA detection.
- Utilized alkaline phosphatase-coupled antibody for signal amplification and NBT/BCIP or BM-Purple substrates for color development.
Main Results:
- Successfully demonstrated a method for localizing miRNAs in skin and hair follicle tissue sections.
- Provided a detailed step-by-step protocol applicable to both frozen and paraffin-embedded samples.
- Established a convenient and effective approach for visualizing miRNA spatiotemporal expression.
Conclusions:
- The described ISH protocol offers a valuable tool for studying miRNA function and regulation.
- This method addresses the challenge of analyzing small RNA molecules like miRNAs.
- The protocol will aid researchers in understanding miRNA roles during development and in various tissues.
Related Concept Videos
02:31In-situ Hybridization
In situ hybridization (ISH) is a technique used to detect and localize specific DNA or RNA molecules in cells, tissue, or tissue sections using a labeled probe. The technique was first used in 1969 for the investigation of nucleic acids. It is currently an essential tool in scientific research and clinical settings, especially for diagnostic purposes.
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
Types of probes and labels
A probe is a complementary strand of DNA or RNA that binds to corresponding nucleotide sequences in a cell. Many...
02:07FISH - Fluorescent In-situ Hybridization
Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...