title: A recording chamber for small volume slice electrophysiology.
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.
authors.affiliation: Department of Physiology and Biophysics, School of Medicine, University of Colorado, Aurora, Colorado; anna.dondzillo@ucdenver.edu.
authors.forename: Anna
authors.initials: A
authors.lastname: Dondzillo
authors.affiliation: Department of Biomedical Sciences, National Jewish Health, Denver, Colorado; and.
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Quinn
authors.affiliation: Department of Biomedical Sciences, National Jewish Health, Denver, Colorado; and.
authors.forename: Charmion I
authors.initials: CI
authors.lastname: Cruickshank-Quinn
authors.affiliation: Department of Biomedical Sciences, National Jewish Health, Denver, Colorado; and.
authors.forename: Nichole
authors.initials: N
authors.lastname: Reisdorph
authors.affiliation: Department of Electrical Engineering, College of Engineering and Applied Science, University of Colorado, Denver, Colorado.
authors.forename: Tim C
authors.initials: TC
authors.lastname: Lei
authors.affiliation: Department of Physiology and Biophysics, School of Medicine, University of Colorado, Aurora, Colorado;
authors.forename: Achim
authors.initials: A
authors.lastname: Klug
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology / Q000502:physiology*; D004594:Electrophysiology / Q000295:instrumentation* / Q000379:methods; D005849:Gerbillinae; D009474:Neurons / Q000502:physiology; D018408:Patch-Clamp Techniques / Q000295:instrumentation* / Q000379:methods
keywords: brain slice; in vitro; patch clamp; recording chamber; slice recordings