Development of μ-Low-Flow-Push-Pull Perfusion Probes for Ex Vivo Sampling from Mouse Hippocampal Tissue Slices

ACS Chemical Neuroscience
Marissa R CabayScott A Shippy

Abstract

This work demonstrates a reduced tip μ-low-flow-push-pull perfusion technique for ex vivo sampling of the extracellular space of mouse hippocampal brain slices. Concentric fused-silica capillary probes are pulled by an in-house gravity puller with a butane flame producing probe tips averaging an overall outer diameter of 30.3 ± 8 μm. The 10-30 nL/min perfusion flow rate through the probe generates an average recovery of 90%. Sampling was performed with mouse brain tissue slices to characterize basal neurotransmitter content in this model system. Samples were collected from hippocampal tissue slices at a volume of 200 nL per sample. Sample arginine, histamine, lysine, glycine, glutamate, and aspartate content was quantified by micellar electrokinetic chromatography with LED-induced fluorescence detection. Primary amine content was sampled over several hours to determine evidence for tissue damage and loss of extracellular content from the tissue slice. Overall, all amino acid concentrations trended lower as an effect of time relative to tissue slicing. There were significant concentration decreases seen for histamine, lysine, and aspartate between time points 0-2 and 2-6 h (p < 0.05) relative to tissue slicing. Analysis of avera...Continue Reading

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Citations

Nov 22, 2018·Analytical and Bioanalytical Chemistry·Anis Barmada, Scott A Shippy
Jun 20, 2020·ACS Chemical Neuroscience·Amir H FarajiStephen G Weber

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