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使用多孔多电极阵列进行视网膜记录的分步说明。

Step-by-step instructions for retina recordings with perforated multi electrode arrays.

作者信息

Reinhard Katja, Tikidji-Hamburyan Alexandra, Seitter Hartwig, Idrees Saad, Mutter Marion, Benkner Boris, Münch Thomas A

机构信息

Werner Reichardt Centre for Integrative Neuroscience and Bernstein Center for Computational Neuroscience, University of Tübingen, Tübingen, Germany.

出版信息

PLoS One. 2014 Aug 28;9(8):e106148. doi: 10.1371/journal.pone.0106148. eCollection 2014.

Abstract

Multi-electrode arrays are a state-of-the-art tool in electrophysiology, also in retina research. The output cells of the retina, the retinal ganglion cells, form a monolayer in many species and are well accessible due to their proximity to the inner retinal surface. This structure has allowed the use of multi-electrode arrays for high-throughput, parallel recordings of retinal responses to presented visual stimuli, and has led to significant new insights into retinal organization and function. However, using conventional arrays where electrodes are embedded into a glass or ceramic plate can be associated with three main problems: (1) low signal-to-noise ratio due to poor contact between electrodes and tissue, especially in the case of strongly curved retinas from small animals, e.g. rodents; (2) insufficient oxygen and nutrient supply to cells located on the bottom of the recording chamber; and (3) displacement of the tissue during recordings. Perforated multi-electrode arrays (pMEAs) have been found to alleviate all three issues in brain slice recordings. Over the last years, we have been using such perforated arrays to study light evoked activity in the retinas of various species including mouse, pig, and human. In this article, we provide detailed step-by-step instructions for the use of perforated MEAs to record visual responses from the retina, including spike recordings from retinal ganglion cells and in vitro electroretinograms (ERG). In addition, we provide in-depth technical and methodological troubleshooting information, and show example recordings of good quality as well as examples for the various problems which might be encountered. While our description is based on the specific equipment we use in our own lab, it may also prove useful when establishing retinal MEA recordings with other equipment.

摘要

多电极阵列是电生理学领域的一种先进工具,在视网膜研究中也是如此。视网膜的输出细胞,即视网膜神经节细胞,在许多物种中形成单层结构,并且由于它们靠近视网膜内表面,因此很容易进行检测。这种结构使得多电极阵列能够用于对呈现的视觉刺激进行视网膜反应的高通量、并行记录,并对视网膜的组织和功能有了重要的新认识。然而,使用将电极嵌入玻璃或陶瓷板中的传统阵列可能会带来三个主要问题:(1)由于电极与组织之间接触不良导致信噪比低,特别是在小动物(如啮齿动物)的视网膜强烈弯曲的情况下;(2)记录室底部的细胞氧气和营养供应不足;(3)记录过程中组织移位。已发现穿孔多电极阵列(pMEA)可缓解脑片记录中的所有这三个问题。在过去几年中,我们一直在使用这种穿孔阵列来研究包括小鼠、猪和人类在内的各种物种视网膜中的光诱发活动。在本文中,我们提供了使用穿孔多电极阵列记录视网膜视觉反应的详细分步说明,包括视网膜神经节细胞的尖峰记录和体外视网膜电图(ERG)。此外,我们提供了深入的技术和方法故障排除信息,并展示了高质量的示例记录以及可能遇到的各种问题的示例。虽然我们的描述基于我们自己实验室使用的特定设备,但在使用其他设备进行视网膜多电极阵列记录时也可能会证明是有用的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/474f/4148441/204be9f44cac/pone.0106148.g001.jpg

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