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Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization

Zebrafish Whole Mount High-Resolution Double Fluorescent In Situ Hybridization

Published on: March 25, 2009

Zinc-based fixation for high-sensitivity in situ hybridization: a nonradioactive colorimetric method for the

Electra Stylianopoulou¹, George Skavdis, Maria Grigoriou

¹Department of Molecular Biology and Genetics, Democritus University of Thrace, Alexandroupolis, Greece.

Methods in Molecular Biology (Clifton, N.J.)

|September 15, 2014

Abstract:

Nonradioactive colorimetric in situ hybridization (NoRISH) has been widely applied to analyze gene expression at the single-cell level. Zinc fixation is time efficient and provides excellent tissue morphology. Furthermore, it improves the preservation of the RNA, facilitating the detection of rare transcripts or the identification of expressing cells scattered within a tissue. Here we present a rapid, highly sensitive NoRISH method that uses a zinc-salt-based fixative and is especially suitable for the study of genes expressed at low levels and/or in a small number of cells within a structure.

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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...

FISH - Fluorescent In-situ Hybridization

FISH - 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,...

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