Department of Cell Biology, State University of New York Downstate Health Sciences University, Brooklyn, NY, USA.
Neural and Behavioral Science Graduate Program, State University of New York Downstate Health Sciences University, Brooklyn, NY, USA.
Sci Rep. 2023 Nov 16;13(1):20005. doi: 10.1038/s41598-023-45949-9.
Acute brain slices are a common and useful preparation in experimental neuroscience. A wide range of incubation chambers for brain slices exists but only a few are designed with very low volumes of the bath solution in mind. Such chambers are necessary when high-cost chemicals are to be added to the solution or when small amounts of substances released by the slice are to be collected for analysis. The principal challenge in designing a very low-volume incubation chamber is maintaining good oxygenation and flow without mechanically disturbing or damaging the slices. We designed and validated BubbleDrive, a 3D-printed incubation chamber with a minimum volume of 1.5 mL which can hold up to three coronal mouse slices from one hemisphere. It employs the carbogen gas bubbles to drive the flow circulation in a consistent and reproducible manner, and without disturbing the brain slices. The BubbleDrive design and construction were successfully validated by comparison to a conventional large-volume incubation chamber in several experimental designs involving measurements of extracellular diffusion parameters, the electrophysiology of neuronal and astrocytic networks, and the effectiveness of slice incubation with hyaluronidase enzyme.
急性脑片是实验神经科学中常用且有用的一种制备方法。有各种各样的脑片孵育室，但只有少数设计考虑到浴液的体积非常小。当需要向溶液中添加高成本化学品或需要收集切片释放的少量物质进行分析时，就需要使用这种小体积孵育室。设计一个非常小体积的孵育室的主要挑战是在不机械干扰或损坏切片的情况下保持良好的氧合和流动。我们设计并验证了 BubbleDrive，这是一种 3D 打印孵育室，最小体积为 1.5 毫升，可以容纳来自一个半球的三个冠状老鼠切片。它利用碳化气体气泡以一致且可重复的方式驱动流动循环，而不会干扰脑切片。通过与传统的大容量孵育室在涉及测量细胞外扩散参数、神经元和星形胶质细胞网络的电生理学以及透明质酸酶孵育切片的有效性的几个实验设计进行比较，成功验证了 BubbleDrive 的设计和构建。
