Related Experiment Video
Updated: May 27, 2026
17:38
Radioactive in situ Hybridization for Detecting Diverse Gene Expression Patterns in Tissue
Published on: April 27, 2012
Studying gene expression in bone by in situ hybridization
Ina Kramer1, Rishard Salie, Mira Susa
1Musculoskeletal Disease Department, Novartis Institutes for BioMedical Research, Basel, Switzerland. ina.kramer@novartis.com
Methods in Molecular Biology (Clifton, N.J.)
|December 2, 2011
Summary
This study presents a nonradioactive in situ hybridization method to detect mRNA in mouse bone sections. The technique utilizes the CryoJane Tape-Transfer System and digoxigenin-labeled riboprobes for accurate transcript localization.
Area of Science:
- Molecular Biology
- Histology
- Biochemistry
Background:
- In situ hybridization (ISH) is crucial for detecting specific nucleic acid sequences within tissue samples.
- Nonradioactive detection methods are preferred for safety and ease of handling compared to radioactive probes.
- Analyzing mRNA expression in bone tissue is vital for understanding skeletal development and disease.
Purpose of the Study:
- To describe a novel method for nonradioactive in situ hybridization.
- To enable the detection of mRNA transcripts in cryostat sections of mouse bone.
- To optimize the use of the CryoJane Tape-Transfer System for bone tissue analysis.
Main Methods:
- Development of a nonradioactive in situ hybridization protocol.
- Application of the CryoJane Tape-Transfer System for preparing cryostat sections.
- Utilizing digoxigenin (DIG)-labeled riboprobes for mRNA detection.
Main Results:
- Successful detection of specific mRNA transcripts in mouse bone cryosections.
- Demonstration of the efficacy of the CryoJane Tape-Transfer System in preserving tissue integrity for ISH.
- Validation of digoxigenin-labeled riboprobes for sensitive and specific transcript visualization.
Conclusions:
- The described method provides a robust approach for nonradioactive mRNA detection in mouse bone.
- This technique facilitates the study of gene expression patterns in skeletal tissues.
- The optimized protocol enhances the reliability and accessibility of in situ hybridization for bone research.
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,...