发布: 2019年03月05日第9卷第5期 DOI: 10.21769/BioProtoc.3187 浏览次数: 5355
评审: Jackeline Moraes MalheirosAnonymous reviewer(s)
Abstract
Acute cerebellar slices are widely used among neuroscientists to study the properties of excitatory and inhibitory synaptic transmission as well as intracellular signaling pathways involved in their regulation in cerebellum. The cerebellar cortex presents a well-organized circuitry, and several neuronal pathways can be stimulated and recorded reliably in acute cerebellar slices. A widely used acute cerebellar slice preparation technique was adapted from Edwards’ thin slice preparation method published in 1989 (Edwards et al., 1989). Most of the acute cerebellar slice preparation techniques use a vibrating microtome for slicing freshly dissected cerebellum from various animal species. Here we introduce a simpler method, which uses a tissue chopper to quickly prepare acute sagittal cerebellar slices from rodents. Cerebellum is dissected from the whole brain and sliced with a tissue chopper into 200-400 µm thick slices. Slices are allowed to recover in oxygenated aCSF at 37 °C for 1-2 h. Slices can then be used for electrophysiology or other types of experimentation. This method can be used to prepare cerebellar slices from mouse or rat aged from postnatal day 7 to 2 years. The preparation is faster and easier than other methods and provides a more versatile diversity of applications.
Keywords: Acute cerebellar slice (小脑快速切片)Background
Acute cerebellar slice preparations, like many other current brain slice preparation techniques, originated from Edwards’ thin slice preparation method published in 1989 (Edwards et al., 1989). In general, cerebellum was quickly dissected and immersed in Ca2+-free aCSF and glued to the stage of a vibratome. After slicing, slices were recovered in regular aCSF for 1-2 h before use (Llano et al., 1991a and 1991b; Kano and Konnerth, 1992). Here we introduce a method adapted from the acute hippocampal slice preparation method used in our laboratory. Freshly dissected cerebellum is rapidly sliced (around 10 s) on a McIlwain tissue chopper without the need for glue or oxygenation during slicing. The slices prepared using this method are as healthy as the ones prepared with a vibratome and can be used for electrophysiology or a variety of other manipulations.
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版权信息
© 2019 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Wang, Y. and Baudry, M. (2019). Acute Cerebellar Slice Preparation Using a Tissue Chopper. Bio-protocol 9(5): e3187. DOI: 10.21769/BioProtoc.3187.
分类
神经科学 > 基础技术 > 快速制片
细胞生物学 > 组织分析 > 组织分离
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