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Development of μ-Low-Flow-Push-Pull Perfusion Probes for Ex Vivo Sampling from Mouse Hippocampal Tissue Slices
Marissa R Cabay1, Alyssa McRay1, David E Featherstone1
1Department of Chemistry, ‡Department of Biological Sciences, and §Laboratory of Integrative Neuroscience, University of Illinois at Chicago , Chicago, Illinois 60607, United States.
ACS Chemical Neuroscience
|October 28, 2017
Summary
This study presents a new micro-perfusion technique for sampling extracellular brain fluid in mouse hippocampal slices. The method uses a smaller probe tip, achieving high recovery rates and enabling neurotransmitter analysis over time.
Area of Science:
- Neuroscience
- Biochemistry
- Analytical Chemistry
Background:
- Accurate sampling of the brain's extracellular space is crucial for understanding neurotransmitter dynamics.
- Previous techniques faced limitations in sensitivity and invasiveness.
- Developing minimally invasive methods is key for studying brain tissue ex vivo.
Purpose of the Study:
- To demonstrate a reduced tip micro-low-flow-push-pull perfusion technique for ex vivo sampling.
- To characterize basal neurotransmitter content in mouse hippocampal slices.
- To assess the impact of sampling time on extracellular content and probe invasiveness.
Main Methods:
- Fabrication of concentric fused-silica capillary probes with reduced tip outer diameter (30.3 ± 8 μm).
- Utilizing a 10-30 nL/min perfusion flow rate for high recovery (average 90%).
- Quantification of amino acids (arginine, histamine, lysine, glycine, glutamate, aspartate) using micellar electrokinetic chromatography with LED-induced fluorescence detection.
Main Results:
- Amino acid concentrations generally decreased over several hours post-slicing.
- Significant decreases in histamine, lysine, and aspartate were observed between 0-2 and 2-6 hours (p < 0.05).
- No significant probe-insertion-related amino acid changes were detected, indicating minimal tissue damage.
Conclusions:
- The reduced tip probe technique is effective for collecting extracellular content from thin brain slices.
- The method allows for reliable neurotransmitter analysis over extended sampling periods.
- This technique offers an improved approach for studying extracellular biochemistry in brain tissue models.
