title: An aerator for brain slice experiments in individual cell culture plate wells.
abstract: BACKGROUND
  Ex vivo acute living brain slices are a broadly employed and powerful experimental preparation. Most new technology regarding this tissue has involved the chamber used when performing electrophysiological experiments. Alternatively we instead focus on the creation of a simple, versatile aerator designed to allow maintenance and manipulation of acute brain slices and potentially other tissue in a multi-well cell culture plate.
  
  
  METHODS
  Here we present an easily manufactured aerator designed to fit into a 24-well cell culture plate. It features a nylon mesh and a single microhole to enable gas delivery without compromising tissue stability. The aerator is designed to be individually controlled, allowing both high throughput and single well experiments.
  
  
  RESULTS
  The aerator was validated by testing material leach, dissolved oxygen delivery, brain slice viability and neuronal electrophysiology. Example experiments are also presented, including a test of whether β1-adrenergic receptor activation regulates gene expression in ex vivo dorsal striatum using qPCR.
  
  
  METHODS
  Key differences include enhanced control over gas delivery to individual wells containing brain slices, decreased necessary volume, a sample restraint to reduce movement artifacts, the potential to be sterilized, the avoidance of materials that absorb water and small biological molecules, minimal production costs, and increased experimental throughput.
  
  
  CONCLUSIONS
  This new aerator is of high utility and will be useful for experiments involving brain slices and other potentially tissue samples in 24-well cell culture plates.
authors.affiliation: Department of Biological Sciences, North Carolina State University, Raleigh, NC, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Dorris
authors.affiliation: Department of Biological Sciences, North Carolina State University, Raleigh, NC, USA.
authors.forename: Caitlin A
authors.initials: CA
authors.lastname: Hauser
authors.affiliation: Department of Biological Sciences, North Carolina State University, Raleigh, NC, USA.
authors.forename: Caitlin E
authors.initials: CE
authors.lastname: Minnehan
authors.affiliation: Department of Biological Sciences, North Carolina State University, Raleigh, NC, USA; W.M. Keck Center for Behavioral Biology, Center for Human Health and the Environment, and Center for Comparative Medicine and Translational Research, North Carolina State University, Raleigh, NC, USA. Electronic address: jemeitze@ncsu.edu.
authors.forename: John
authors.initials: J
authors.lastname: Meitzen
mesh_terms: D000818:Animals; D001921:Brain / Q000187:drug effects / Q000502:physiology*; D018929:Cell Culture Techniques / Q000295:instrumentation*; D004867:Equipment Design; D054769:G-Protein-Coupled Receptor Kinase 2 / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects / Q000502:physiology; D007150:Immunohistochemistry; D009474:Neurons / Q000187:drug effects / Q000502:physiology; D009924:Organ Culture Techniques / Q000295:instrumentation*; D010100:Oxygen / Q000008:administration & dosage; D018408:Patch-Clamp Techniques / Q000295:instrumentation; D012333:RNA, Messenger / Q000378:metabolism; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D013057:Spectrum Analysis
chemical_list: D012333:RNA, Messenger; C496569:Grk2 protein, rat; D054769:G-Protein-Coupled Receptor Kinase 2; D010100:Oxygen
keywords: 24-Well culture plate; Aerator; Brain slices; Bubbler; Caudate putamen; Dorsal striatum; Oxygenation; β-Adrenergic receptor