Related Experiment Video
Updated: Feb 6, 2026
10:01
Probing mRNA Kinetics in Space and Time in Escherichia coli using Two-Color Single-Molecule Fluorescence In Situ Hybridization
Published on: July 30, 2020
7.9K
Real-Time Monitoring of Fluorescence in Situ Hybridization Kinetics
Nadya Ostromohov1,2, Deborah Huber1, Moran Bercovici2
1IBM Research-Zurich , Säumerstrasse 4 , 8803 Rüschlikon , Zurich , Switzerland.
Analytical Chemistry
|August 21, 2018
Summary
This study introduces a new method for real-time monitoring of fluorescence in situ hybridization (FISH) kinetics. The technique uses a microfluidic probe to analyze factors influencing FISH assay efficiency.
Area of Science:
- Molecular Biology
- Biotechnology
- Analytical Chemistry
Background:
- Fluorescence in situ hybridization (FISH) is a powerful technique for visualizing specific DNA sequences.
- Real-time kinetic analysis of FISH can provide deeper insights into hybridization efficiency and assay optimization.
- Current methods may lack the speed and precision for dynamic monitoring of FISH processes.
Purpose of the Study:
- To develop and validate a novel method for real-time monitoring and kinetic analysis of FISH.
- To investigate the influence of various experimental parameters on FISH kinetics.
- To facilitate the rational design and optimization of FISH assays.
Main Methods:
- Implementation of a vertical microfluidic probe with rapid switching capabilities.
- Real-time monitoring of FISH signal during imaging buffer wash to remove unbound probes.
- Application of the method to a model system using centromeric probes (Cen17).
Main Results:
- Demonstrated applicability for characterizing FISH kinetics under varying conditions.
- Quantified the effects of probe concentration, formamide, dextran sulfate, and ionic strength.
- Provided insights into the dynamic processes governing in situ hybridization.
Conclusions:
- The developed method enables precise, real-time kinetic analysis of FISH.
- It offers a valuable tool for understanding and optimizing FISH assay parameters.
- Facilitates the development of improved FISH-based diagnostic and research tools.
Related Concept Videos
02:07FISH - Fluorescent In-situ Hybridization
24.5K
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,...
24.5K
02:31In-situ Hybridization
10.6K
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...
10.6K
02:57Real Time RT-PCR
65.3K
Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...
The real-time quantification of the number of amplified products is...
65.3K
00:23Kinetic Energy
43.5K
Kinetic energy is the ability of an object in motion to do work or enact change. It can take on many forms. For instance, water flowing down a waterfall has kinetic energy. In biological systems, particles of light travel and are absorbed by plants to create chemical energy. Animals consume the chemical energy and give off molecules that carry their scent through the air. They also generate kinetic energy when they run away from predators. Entire systems also possess kinetic energy, like the...
43.5K
01:19Enzyme Kinetics
104.2K
Enzymes speed up reactions by lowering the activation energy of the reactants. The speed at which the enzyme turns reactants into products is called the rate of reaction. Several factors impact the rate of reaction, including the number of available reactants. Enzyme kinetics is the study of how an enzyme changes the rate of a reaction.
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...
Scientists typically study enzyme kinetics with a fixed amount of enzyme in the controlled environment of a test tube. When more reactant, or substrate, is...
104.2K
02:29Hybrid Zones
21.9K
Hybrid zones are narrow regions where two closely related species interact, mate, and produce hybrids. Relative to either parent species, hybrids may possess distinct phenotypic or genetic differences that impact their survival and reproductive success. The genetic variances introduced by hybridization influence species diversity and speciation processes within the hybrid zone.
21.9K