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1.
J Neural Eng ; 16(6): 066021, 2019 10 29.
Article in English | MEDLINE | ID: mdl-31216526

ABSTRACT

OBJECTIVE: Electrode arrays for chronic implantation in the brain are a critical technology in both neuroscience and medicine. Recently, flexible, thin-film polymer electrode arrays have shown promise in facilitating stable, single-unit recordings spanning months in rats. While array flexibility enhances integration with neural tissue, it also requires removal of the dura mater, the tough membrane surrounding the brain, and temporary bracing to penetrate the brain parenchyma. Durotomy increases brain swelling, vascular damage, and surgical time. Insertion using a bracing shuttle results in additional vascular damage and brain compression, which increase with device diameter; while a higher-diameter shuttle will have a higher critical load and more likely penetrate dura, it will damage more brain parenchyma and vasculature. One way to penetrate the intact dura and limit tissue compression without increasing shuttle diameter is to reduce the force required for insertion by sharpening the shuttle tip. APPROACH: We describe a novel design and fabrication process to create silicon insertion shuttles that are sharp in three dimensions and can penetrate rat dura, for faster, easier, and less damaging implantation of polymer arrays. Sharpened profiles are obtained by reflowing patterned photoresist, then transferring its sloped profile to silicon with dry etches. MAIN RESULTS: We demonstrate that sharpened shuttles can reliably implant polymer probes through dura to yield high quality single unit and local field potential recordings for at least 95 days. On insertion directly through dura, tissue compression is minimal. SIGNIFICANCE: This is the first demonstration of a rat dural-penetrating array for chronic recording. This device obviates the need for a durotomy, reducing surgical time and risk of damage to the blood-brain barrier. This is an improvement to state-of-the-art flexible polymer electrode arrays that facilitates their implantation, particularly in multi-site recording experiments. This sharpening process can also be integrated into silicon electrode array fabrication.


Subject(s)
Brain/physiology , Dura Mater/physiology , Electrodes, Implanted , Equipment Design/methods , Microtechnology/methods , Silicon , Animals , Biocompatible Materials , Equipment Design/instrumentation , Male , Microelectrodes , Microtechnology/instrumentation , Rats , Rats, Long-Evans
2.
Lab Chip ; 18(15): 2291-2299, 2018 07 24.
Article in English | MEDLINE | ID: mdl-29987290

ABSTRACT

A nucleic acid amplification-free, optics-free platform has been demonstrated for sequence-specific detection of Escherichia coli (E. coli) 16S rRNA at 1 aM (10-18 M) against a 106-fold (1 pM) background of Pseudomonas putida (P. putida) RNA. This work was driven by the need for simple, rapid, and low cost means for species-specific bacterial detection at low concentration. Our simple, conductometric sensing device functioned by detecting blockage of a nanopore fabricated in a sub-micron-thick glass membrane. Upon sequence-specific binding of target 16S rRNA, otherwise charge-neutral, PNA oligonucleotide probe-polystyrene bead conjugates become electrophoretically mobile and are driven to the glass nanopore of lesser diameter, which is blocked, thereby generating a large, sustained and readily observable step decrease in ionic current. No false positive signals were observed with P. putida RNA when this device was configured to detect E. coli 16S rRNA. Also, when a universal PNA probe complementary to the 16S rRNA of both E. coli and P. putida was conjugated to beads, a positive response to rRNA of both bacterial species was observed. Finally, the device readily detected E. coli at 10 CFU mL-1 in a 1 mL sample, also against a million-fold background of viable P. putida. These results suggest that this new device may serve as the basis for small, portable, low power, and low-cost systems for rapid detection of specific bacterial species in clinical samples, food, and water.


Subject(s)
Biosensing Techniques/methods , Escherichia coli/isolation & purification , Pseudomonas putida/isolation & purification , RNA, Ribosomal, 16S/analysis , RNA, Ribosomal, 16S/genetics , Base Sequence , Biosensing Techniques/economics , Costs and Cost Analysis , Escherichia coli/genetics , Limit of Detection , Microspheres , Nucleic Acid Hybridization , Peptide Nucleic Acids/chemistry , Pseudomonas putida/genetics , RNA, Ribosomal, 16S/chemistry , Time Factors
4.
Sci Rep ; 5: 12511, 2015 Jul 27.
Article in English | MEDLINE | ID: mdl-26212790

ABSTRACT

Environmental stimuli have the ability to generate specific representations of the rewards they predict and in so doing alter the selection and performance of reward-seeking actions. The basolateral amygdala participates in this process, but precisely how is unknown. To rectify this, we monitored, in near-real time, basolateral amygdala glutamate concentration changes during a test of the ability of reward-predictive cues to influence reward-seeking actions (Pavlovian-instrumental transfer). Glutamate concentration was found to be transiently elevated around instrumental reward seeking. During the Pavlovian-instrumental transfer test these glutamate transients were time-locked to and correlated with only those actions invigorated by outcome-specific motivational information provided by the reward-predictive stimulus (i.e., actions earning the same specific outcome as predicted by the presented CS). In addition, basolateral amygdala AMPA, but not NMDA glutamate receptor inactivation abolished the selective excitatory influence of reward-predictive cues over reward seeking. These data support [corrected] the hypothesis that transient glutamate release in the BLA can encode the outcome-specific motivational information provided by reward-predictive stimuli.


Subject(s)
Amygdala/metabolism , Anticipation, Psychological/physiology , Conditioning, Operant/physiology , Glutamic Acid/metabolism , Reward , Adaptation, Physiological/physiology , Animals , Choice Behavior/physiology , Cues , Extinction, Psychological/physiology , Male , Neurotransmitter Agents/metabolism , Rats , Rats, Long-Evans
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