Dept. of Pharmacology, University of Oxford, Mansfield Road, Oxford, United Kingdom.
J Neurosci Methods. 2011 Jan 30;195(1):15-23. doi: 10.1016/j.jneumeth.2010.10.024. Epub 2010 Nov 12.
In vitro brain slice electrophysiology is a powerful and highly successful technique where a thin slice is cut from the brain and kept alive artificially in a recording chamber. The design of this recording chamber is pivotal to the success and the quality of such experiments. Most often one of two types of chambers is used today, the interface chamber or the submerged chamber. These chambers, however, have the disadvantage that they are limited in either their experimental or their physiological properties respectively. Here we present a new working principle for an in vitro chamber design which aims at combining the advantages of the classical designs whilst overcoming their disadvantages. This is achieved by using a semipermeable membrane on which the slice is placed. The membrane allows for a fast flow of artificial cerebrospinal fluid of up to at least 17 ml/min. Due to a Bernoulli effect, this high speed flow also causes a 64% increase in flow of solution across the membrane on which the slice rests. The fact that the membrane is transparent introduces the possibility of wide field inverted optical imaging to brain slice electrophysiology. The utility of this setup was demonstrated in the recording of local field potential, single cell and voltage sensitive dye imaging data simultaneously from an area smaller then 1/8mm(2). The combination of all these features in the membrane chamber make it a versatile and promising device for many current and future in vitro applications, especially in the regard to optical imaging.
体外脑片电生理学是一种强大且非常成功的技术，它将脑的薄片从大脑中取出并在记录室中人工维持生命。该记录室的设计对实验的成功和质量至关重要。今天，最常用的是两种类型的腔室之一，即界面腔室或淹没腔室。然而，这些腔室各自具有实验或生理特性受限的缺点。在这里，我们提出了一种新的体外腔室设计原理，旨在结合经典设计的优点，同时克服它们的缺点。这是通过在薄片上放置半透膜来实现的。该膜允许人工脑脊液以高达至少 17ml/min 的快速流速通过。由于伯努利效应，这种高速流动也会导致在薄片所在的膜上的溶液流量增加 64%。膜是透明的这一事实为脑片电生理学引入了广角倒置光学成像的可能性。该设置的实用性在记录局部场电位、单细胞和电压敏感染料成像数据方面得到了证明，同时记录的区域小于 1/8mm(2)。这些特征在膜腔中的结合使其成为许多当前和未来体外应用的多功能且有前途的设备，特别是在光学成像方面。
