Related Experiment Video
Updated: Feb 11, 2026
09:05
Multiplex Detection of Gene Expression in the Intact Drosophila Brain Using Expansion-Assisted Iterative Fluorescence In Situ Hybridization
Published on: May 2, 2025
1.7K
itFISH: Enhanced Staining by Iterative Fluorescent In Situ Hybridization
Richard H Row1, Benjamin L Martin1
1Department of Biochemistry and Cell Biology, Stony Brook University , Stony Brook, New York.
Zebrafish
|March 21, 2017
Summary
This study enhances fluorescent in situ hybridization (FISH) for zebrafish research. The new method significantly improves detection of weakly expressed genes by iterating antibody staining and development steps.
Area of Science:
- Developmental Biology
- Molecular Biology
- Genetics
Background:
- Fluorescent in situ hybridization (FISH) is crucial for analyzing gene expression in zebrafish embryos.
- Tyramide signal amplification is a common, cost-effective FISH method.
- Detecting weakly expressed genes remains a challenge with current FISH techniques.
Purpose of the Study:
- To adapt a planarian-based method for enhanced FISH signal in zebrafish.
- To improve the detection sensitivity of FISH for low-abundance transcripts.
- To overcome limitations of standard FISH for weakly expressed genes.
Main Methods:
- Adapted a signal amplification protocol from planarian research for zebrafish.
- Iterated antibody staining and enzymatic development steps.
- Applied the enhanced method to detect previously undetectable gene expression.
Main Results:
- Achieved dramatic signal enhancement for previously weak FISH signals.
- Successfully detected gene expression from probes that were previously too weak to visualize.
- Demonstrated a robust method for sensitive gene expression analysis in zebrafish.
Conclusions:
- The adapted iterative staining method significantly enhances FISH sensitivity in zebrafish.
- This technique enables robust detection of weakly expressed genes, expanding research capabilities.
- The enhanced FISH approach offers a powerful tool for studying gene expression patterns in developmental biology.
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
02:29Hybrid Zones
22.0K
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.
22.0K
03:24Hybridization of Atomic Orbitals I
67.7K
The mathematical expression known as the wave function, ψ, contains information about each orbital and the wavelike properties of electrons in an isolated atom. When atoms are bound together in a molecule, the wave functions combine to produce new mathematical descriptions that have different shapes. This process of combining the wave functions for atomic orbitals is called hybridization and is mathematically accomplished by the linear combination of atomic orbitals. The new orbitals that...
67.7K
03:35Hybridization of Atomic Orbitals II
49.2K
sp3d and sp3d 2 Hybridization
49.2K
03:00Self-Evaluation: Self-Enhancement and Self-Verification
5.8K
Social psychologists have documented that feeling good about ourselves and maintaining positive self-esteem is a powerful motivator of human behavior (Tavris & Aronson, 2008). In the United States, members of the predominant culture typically think very highly of themselves and view themselves as good people who are above average on many desirable traits (Ehrlinger, Gilovich, & Ross, 2005). Often, our behavior, attitudes, and beliefs are affected when we experience a threat to our...
5.8K