An aerator for brain slice experiments in individual cell culture plate wells

• Published in: Journal of neuroscience methods 2014-12, Vol.238, p.1-10
• Main Authors: Dorris, David M., Hauser, Caitlin A., Minnehan, Caitlin E., Meitzen, John
• Format: Article
• Language: English
• Subjects: 24-Well culture plate; Aerator; Animals; Brain - drug effects; Brain - physiology; Brain slices; Bubbler; Caudate putamen; Cell Culture Techniques - instrumentation; Dorsal striatum; Equipment Design; G-Protein-Coupled Receptor Kinase 2 - metabolism; Gene Expression - drug effects; Gene Expression - physiology; Immunohistochemistry; Neurons - drug effects; Neurons - physiology; Organ Culture Techniques - instrumentation; Oxygen - administration & dosage; Oxygenation; Patch-Clamp Techniques - instrumentation; Rats, Sprague-Dawley; Real-Time Polymerase Chain Reaction; RNA, Messenger - metabolism; Spectrum Analysis; β-Adrenergic receptor
• ISSN: 0165-0270; 1872-678X
• DOI: 10.1016/j.jneumeth.2014.09.017

•We have developed a new aerator designed to fit into a single well of a standard 24-well cell culture plate.•The aerator keeps brain slices viable and stationary, and is inexpensive to produce.•The aerator enables individual manipulation of living acute brain slices or potentially other tissues in low solution volumes. 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. 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. 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. 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. 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.