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

作者：Dorris David M; Hauser Caitlin A; Minnehan Caitlin E; Meitzen John^*

来源：Journal of Neuroscience Methods, 2014, 238: 1-10.

DOI：10.1016/j.jneumeth.2014.09.017

• Brain slices
• Aerator
• Bubbler
• 24-Well culture plate
• Oxygenation
• Dorsal striatum
• beta-Adrenergic receptor
• Caudate putamen

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. %26lt;br%26gt;New method: 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. %26lt;br%26gt;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 beta 1-adrenergic receptor activation regulates gene expression in ex vivo dorsal striatum using qPCR. %26lt;br%26gt;Comparison with existing 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. %26lt;br%26gt;Conclusion: 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.

• 出版日期2014-12-30