Int J Oral Biol.  2015 Dec;40(4):183-187. 10.11620/IJOB.2015.40.4.183.

The Influence of Glutaraldehyde Concentration on Electron Microscopic Multiple Immunostaining

• Bae JS
• Yeo EJ
• Bae YC

• KMID: 2133222
• DOI: http://doi.org/10.11620/IJOB.2015.40.4.183

Abstract

The present study was aimed to evaluate the influence of glutaraldehyde (GA) concentration on multiple electron microscopic (EM) immunostaining using pre-embedding peroxidase and post-embedding immunogold method. Influence of various concentrations of GA included in the fixative on immuoreactivity was assessed in the multiple immunostaining using antisera against anti-transient receptor potential vanilloid 1 (TRPV1) for peroxidase staining and anti-GABA for immunogold labeling in the rat trigeminal caudal nucleus. Anti-TRPV1 antiserum had specificity in pre-embedding peroxidase staining when tissues were fixed with fixative containing paraformaldehyde (PFA) alone. Immunoreactivity for TRPV1 was specific in tissues fixed with fixative containing 0.5% GA at both perfusion and postfixation steps, though the immunoreactivity was weaker than in tissues fixed with fixative containing PFA alone. Tissues fixed with fixative containing 0.5% GA at the perfusion and postfixation steps showed specific immunogold staining for GABA. The results of the present study indicate that GA concentration is critical for immunoreactivity to antigens such as TRPV1 and GABA. This study also suggests that the appropriate GA concentration is 0.5% for multiple immunostaining with peroxidase labeling for TRPV1 and immunogold labeling for GABA.

Keyword

fixation; glutaraldehyde concentration; immunoreactivity; multiple immunostaining; electron microscopy

MeSH Terms

Animals
gamma-Aminobutyric Acid
Glutaral*
Immune Sera
Microscopy, Electron
Perfusion
Peroxidase
Rats
Sensitivity and Specificity
Trigeminal Caudal Nucleus
Glutaral
Immune Sera
Peroxidase
gamma-Aminobutyric Acid