Fluorescence in situ hybridization (FISH) on tissue cryosections

Area of Science:

• Cell Biology
• Molecular Biology
• Genomics

Background:

• Fluorescence in situ hybridization (FISH) has advanced understanding of nuclear organization.
• Most FISH studies use cultured cells, which may not reflect in vivo nuclear processes.
• Nuclear organization and function can differ significantly across tissues and in whole organisms compared to cell cultures.

Purpose of the Study:

• To develop and present a robust FISH protocol for analyzing nuclear organization in tissue sections.
• To address the methodological challenges hindering the application of FISH in tissue samples.
• To facilitate a better understanding of nuclear processes in ex vivo organismal material.

Main Methods:

• The study details a FISH protocol optimized for cryosections of various tissues.
• The protocol is based on extensive practical experience with different probes and tissue types.
• Emphasis is placed on overcoming common methodological hurdles in tissue-based FISH.

Main Results:

• The presented protocol enables successful hybridization of probes on tissue cryosections.
• It provides a viable method for studying nuclear organization in intact tissues.
• The methodology is adaptable for diverse tissues and biological questions.

Conclusions:

• FISH on tissue sections is crucial for understanding in vivo nuclear organization.
• The developed protocol offers a standardized and effective approach for researchers.
• This method will enhance studies on nuclear function in the context of whole organisms and different tissues.