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.

Study Duration

Not specified

Participants

20 adult female cats

Evidence Level

Not specified

Key Findings

1
Triton™ decreases the time required for long-distance transport of FG from the spinal cord to the motor cortex by >4 fold.
2
Higher FG concentrations lead to a greater amount of tissue damage at the injection site.
3
FG 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

The study introduces a method to enhance FG uptake using Triton™, reducing FG-related tissue damage while maintaining effective quantification. Triton™ enhances FG uptake, reduces required FG, and substantially decreases tracing time, limiting FG-induced motor deficits. Detection challenges from small FG concentration are resolved using FG anti-serum and chromogen reactions.

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

1
Specific cell death markers or degeneration stains were not used to assess FG-induced damage.
2
The enhanced tracing speed with Triton™ might potentially trace diseased, non-functional axons.
3
The FG fluorescence is very low in these long distance targets compared to that typically reported in studies with rodents.

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