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Enhancing Fluorogold-based neural tract tracing
S E Mondello1, S C Jefferson2, W A O'Steen3
1Department of Rehabilitation Medicine, University of Washington, Seattle, WA 98195, United States.
Journal of Neuroscience Methods
|June 12, 2016
Summary
This study introduces a method using Triton™ to enhance Fluorogold (FG) neural tracing. The new technique reduces FG toxicity and tracing time, minimizing motor deficits and improving detection sensitivity.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Fluorogold (FG) is a widely used retrograde tracer for nervous system pathways.
- However, FG exhibits neurotoxicity, causing injection site damage and motor deficits.
Purpose of the Study:
- To describe a novel method for enhancing FG uptake.
- To present a procedure for minimizing FG-related tissue damage and improving quantification.
Main Methods:
- Utilized Triton™ to enhance FG uptake and transport.
- Employed FG antiserum with an ABC chromogen reaction for improved fluorescence detection.
- Investigated long-distance tracing from thoracic spinal cord to motor cortex.
Main Results:
- Triton™ enhanced FG uptake and reduced required FG amounts and tracing time by over 4-fold for distances greater than 10 inches.
- Reduced FG concentrations and volumes minimized tissue damage and motor deficits.
- FG antiserum and ABC chromogen reaction provided stable, detectable fluorescence.
Conclusions:
- Triton™ enhances FG neural tracing by improving uptake and reducing toxicity and tracing time.
- The method effectively minimizes FG-induced tissue damage and motor deficits.
- Improved detection methods ensure reliable quantification despite low FG concentrations.
