| Literature DB >> 26203105 |
Anna Dondzillo1, Kevin D Quinn2, Charmion I Cruickshank-Quinn2, Nichole Reisdorph2, Tim C Lei3, Achim Klug4.
Abstract
Electrophysiological recordings from brain slices are typically performed in small recording chambers that allow for the superfusion of the tissue with artificial extracellular solution (ECS), while the chamber holding the tissue is mounted in the optical path of a microscope to image neurons in the tissue. ECS itself is inexpensive, and thus superfusion rates and volumes of ECS consumed during an experiment using standard ECS are not critical. However, some experiments require the addition of expensive pharmacological agents or other chemical compounds to the ECS, creating a need to build superfusion systems that operate on small volumes while still delivering appropriate amounts of oxygen and other nutrients to the tissue. We developed a closed circulation tissue chamber for slice recordings that operates with small volumes of bath solution in the range of 1.0 to 2.6 ml and a constant oxygen/carbon dioxide delivery to the solution in the bath. In our chamber, the ECS is oxygenated and recirculated directly in the recording chamber, eliminating the need for tubes and external bottles/containers to recirculate and bubble ECS and greatly reducing the total ECS volume required for superfusion. At the same time, the efficiency of tissue oxygenation and health of the section are comparable to standard superfusion methods. We also determined that the small volume of ECS contains a sufficient amount of nutrients to support the health of a standard brain slice for several hours without concern for either depletion of nutrients or accumulation of waste products.Entities:
Keywords: brain slice; in vitro; patch clamp; recording chamber; slice recordings
Mesh:
Year: 2015 PMID: 26203105 PMCID: PMC4588903 DOI: 10.1152/jn.00289.2014
Source DB: PubMed Journal: J Neurophysiol ISSN: 0022-3077 Impact factor: 2.714