Buskila Yossi, Breen Paul P, Tapson Jonathan, van Schaik André, Barton Matthew, Morley John W
Bioelectronics and Neuroscience group, The MARCS Institute, University of Western Sydney, Penrith, NSW, Australia.
School of Medicine, University of Western Sydney, Penrith, NSW, Australia.
Sci Rep. 2014 Jun 16;4:5309. doi: 10.1038/srep05309.
The lifespan of an acute brain slice is approximately 6-12 hours, limiting potential experimentation time. We have designed a new recovery incubation system capable of extending their lifespan to more than 36 hours. This system controls the temperature of the incubated artificial cerebral spinal fluid (aCSF) while continuously passing the fluid through a UVC filtration system and simultaneously monitoring temperature and pH. The combination of controlled temperature and UVC filtering maintains bacteria levels in the lag phase and leads to the dramatic extension of the brain slice lifespan. Brain slice viability was validated through electrophysiological recordings as well as live/dead cell assays. This system benefits researchers by monitoring incubation conditions and standardizing this artificial environment. It further provides viable tissue for two experimental days, reducing the time spent preparing brain slices and the number of animals required for research.
急性脑片的寿命约为6至12小时，这限制了潜在的实验时间。我们设计了一种新的恢复孵育系统，能够将其寿命延长至36小时以上。该系统控制孵育的人工脑脊液（aCSF）的温度，同时使液体持续通过紫外线C过滤系统，并同步监测温度和pH值。控制温度和紫外线C过滤相结合，可使细菌水平维持在对数期，从而显著延长脑片的寿命。通过电生理记录以及活/死细胞检测验证了脑片的活力。该系统通过监测孵育条件并使这种人工环境标准化，为研究人员带来了便利。它还能在两个实验日内提供有活力的组织，减少了制备脑片所花费的时间以及研究所需的动物数量。
