Enhancing Fluorogold-based neural tract tracing
J Neurosci Methods, 2016 · DOI: 10.1016/j.jneumeth.2016.06.004 · Published: September 1, 2016
Simple Explanation
Fluorogold (FG) is a tracer used to map nerve pathways, but it can damage tissue and cause motor deficits. Using Triton™ enhances FG uptake, shortens tracing time, and reduces the amount of FG needed, minimizing tissue damage. Although small FG concentrations and injection volumes minimize damage, the resulting fluorescence is difficult to detect, but this can be solved using FG antiserum with a chromogen reaction.
Key Findings
- 1Triton™ decreases the time required for long-distance transport of FG from the spinal cord to the motor cortex by >4 fold.
- 2Higher FG concentrations lead to a greater amount of tissue damage at the injection site.
- 3FG antiserum paired with an ABC chromogen reaction greatly enhances the signal and allows for easy quantification, remaining stable for at least 9 years.
Research Summary
Practical Implications
Improved Neural Tracing
The use of Triton™ enhances the speed and efficiency of neural tracing with Fluorogold.
Reduced Neurotoxicity
Lower concentrations and volumes of Fluorogold, facilitated by Triton™, minimize tissue damage and associated functional deficits.
Enhanced Signal Detection
Immunohistochemical techniques, specifically using an anti-FG antibody and DAB chromogen, amplify the Fluorogold signal for better visualization and analysis.
Study Limitations
- 1Specific cell death markers or degeneration stains were not used to assess FG-induced damage.
- 2The enhanced tracing speed with Triton™ might potentially trace diseased, non-functional axons.
- 3The FG fluorescence is very low in these long distance targets compared to that typically reported in studies with rodents.