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In-situ Hybridization

In-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...

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Related Experiment Video

Updated: Jun 17, 2026

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Single Droplet Digital Polymerase Chain Reaction for Comprehensive and Simultaneous Detection of Mutations in Hotspot Regions

Published on: September 25, 2018

Single-Cell Genotyping of Single-Nucleotide Mutations Using In Situ Allele-Specific Loop-Mediated Isothermal

Zilan Yuan¹, Xinmiao Liu¹, Sha Deng¹

¹College of Biomass Science and Engineering, Healthy Food Evaluation Research Center, Sichuan University, Chengdu 610065, Sichuan, China.

|October 20, 2023

Summary

Allele-specific loop-mediated isothermal amplification (AlleLAMP) enables visualization of single-nucleotide mutations (SNMs) within bacterial cells. This method precisely quantifies antibiotic-resistant bacteria, revealing their survival advantage under preservative stress.

Keywords:

LAMP Salmonella antibiotic resistance single-cell analysis single-nucleotide mutation

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Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH

Single Cell Analysis Of Transcriptionally Active Alleles By Single Molecule FISH

Published on: September 20, 2020

Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

Single-nucleotide mutations (SNMs) are a key driver of bacterial antibiotic resistance.
Visualizing SNMs at the single-cell level is crucial for understanding resistance phenotypes but presents significant challenges.

Purpose of the Study:

To develop a novel method for imaging and quantifying SNMs within bacterial cells in situ.
To investigate the survival advantage of antibiotic-resistant bacteria under preservative stress using single-cell resolution.

Main Methods:

Developed an allele-specific loop-mediated isothermal amplification (LAMP) technique, termed AlleLAMP.
Designed primers with artificial mismatch bases for allele-specific amplification of SNMs.
Applied AlleLAMP for high-gain imaging and precise quantification of mutated quinolone-resistant Salmonella in mixtures.

Main Results:

AlleLAMP successfully imaged bacterial genes with single-nucleotide resolution.
The method accurately quantified mutated quinolone-resistant Salmonella within bacterial populations.
Quinolone-resistant strains demonstrated a survival advantage over sensitive strains under preservative stress.

Conclusions:

AlleLAMP provides a powerful single-cell tool for visualizing and quantifying SNMs.
The study highlights the role of preservative stress in selecting for antibiotic-resistant bacterial strains.
AlleLAMP facilitates the analysis of genotype-phenotype relationships in bacteria.