Preparing glass slides and coverslips for in situ hybridization

Area of Science:

• Molecular Biology
• Histology

Background:

• Precleaned glass slides are suitable for in situ and immunohistochemical techniques.
• In situ hybridization requires RNase inactivation and enhanced tissue adherence to glass slides.

Purpose of the Study:

• To describe methods for coating glass slides with 3-triethoxysilylpropylamine (TESPA) and poly-L-lysine for in situ hybridization.
• To outline the preparation of coverslips for in situ hybridization.

Main Methods:

• Slides are treated with diethyl pyrocarbonate (DEPC) to eliminate RNase activity.
• Glass slides are coated with either TESPA or poly-L-lysine to improve section adhesion.
• Coverslips are prepared using a siliconizing solution.

Main Results:

• TESPA-coated slides offer long-term storage, with adhesion improving post-drying.
• Poly-L-lysine-coated slides ensure immediate section adhesion but require fresh preparation.
• Both methods aim to prevent section detachment during extensive washing procedures.

Conclusions:

• The choice between TESPA and poly-L-lysine depends on specific experimental needs regarding storage and immediate adhesion.
• Proper slide and coverslip preparation is crucial for successful in situ hybridization outcomes.