A multi-channel device for high-density target-selective stimulation and long-term monitoring of cells and subcellular features in C. elegans.
A multi-channel device for high-density target-selective stimulation and long-term monitoring of cells and subcellular features in C. elegans.
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DOI：
10.1039/c4lc00789a
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发表时间：
2014-12-07
期刊：
影响因子：
6.1
通讯作者：
Lu H
中科院分区：
文献类型：
--
作者：
Lee H;Kim SA;Coakley S;Mugno P;Hammarlund M;Hilliard MA;Lu H
选择性细胞消融可用于鉴定多细胞模式生物如秀丽隐杆线虫中的神经元功能。光遗传学工具KillerRed通过以时间和空间精确的方式实现细胞或亚细胞组分的光激活损伤来促进选择性消融。然而，使用KillerRed需要刺激（5分钟-1小时），培养（~24小时）和成像（经常重复）大量的个体动物。目前的手动操作方法受到其耗时、劳动密集型性质以及其麻醉剂使用的限制。为了促进大规模的选择性消融、培养和重复成像，我们开发了一种密集包装的多通道装置，并使用它对表达KillerRed的动物进行高通量神经元消融。在相同位置以高加载效率加载蠕虫的能力使我们能够同时消融多个蠕虫中的选定神经元。我们的设备还可以在KillerRed激活后连续观察动物24小时，并允许动物恢复进行行为测定。我们希望这种多通道设备能够促进神经科学中广泛的长期成像和选择性照明实验。
Selective cell ablation can be used to identify neuronal functions in multicellular model organisms such as Caenorhabditis elegans. The optogenetic tool KillerRed facilitates selective ablation by enabling light-activated damage of cell or subcellular components in a temporally and spatially precise manner. However, the use of KillerRed requires stimulating (5 min-1 hr), culturing (~24 hrs) and imaging (often repeatedly) a large number of individual animals. Current manual manipulation methods are limited by their time-consuming, labor-intensive nature, and their usage of anesthetics. To facilitate large-scale selective ablation, culturing, and repetitive imaging, we developed a densely-packed multi-channel device and used it to perform high-throughput neuronal ablation on KillerRed-expressing animals. The ability to load worms in identical locations with high loading efficiency allows us to ablate selected neurons in multiple worms simultaneously. Our device also enables continuous observation of aminals for 24 hrs following KillerRed activation, and allows the animals to be recovered for behavioural assays. We expect this multi-channel device to facilitate a broad range of long-term imaging and selective illumination experiments in neuroscience.
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影响因子：
--
作者：
Fang-Yen, Christopher;Gabel, Christopher V.;Samuel, Aravinthan D. T.;Bargmann, Cornelia I.;Avery, Leon
通讯作者：
Avery, Leon
影响因子：
2.5
作者：
Lee, Hyewon;Crane, Matthew M.;Lu, Hang
通讯作者：
Lu, Hang
影响因子：
6.1
作者：
Hulme, S. Elizabeth;Shevkoplyas, Sergey S.;Whitesides, George M.
通讯作者：
Whitesides, George M.
影响因子：
6.1
作者：
Chung K;Zhan M;Srinivasan J;Sternberg PW;Gong E;Schroeder FC;Lu H
通讯作者：
Lu H
用于表征个体秀丽隐杆线虫神经毒素诱导反应的液滴微流体
影响因子：
6.1
作者：
Shi, Weiwei;Wen, Hui;Qin, Jianhua
通讯作者：
Qin, Jianhua