A platform for high throughput two-photon-targeted in vivo cellular physiology

高通量双光子靶向体内细胞生理学平台

• 批准号: BB/K001817/1
• 负责人: Simon Schultz
• 金额: $ 53.24万
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FK001817%2F1
• 关键词: platform; throughput; two; photon; targeted

The whole-cell patch clamp recording technique has had much impact in the Life Sciences by allowing the currents in ion channels to recorded: this has been crucial to advances in our understanding of cellular function. Use of the patch-clamp technique in vivo, however, has been difficult, because it essentially has to be carried out "blind", with the only feedback obtained by monitoring the impedance of the micropipette used to make the recording.Recent developments in multiphoton microscopy now allow us to make targeted recordings from cells that have been fluorescently labelled. It has also recently been shown that in vivo patch-clamp recordings can be automated. In this project, we will combine these principles to develop a new approach for whole cell patch clamp electrophysiology. Our platform will allow genetically targeted classes of cells to be visually selected ("point and click") by a human operator, and then automatically patched by a group of robotic micromanipulators capable of obtaining recordings from up to six cells simultaneously. As well as allowing high throughput characterization of cells in vivo for basic scientific or drug discovery purposes, our platform will allow new scientific questions to be asked involving interactions between cells that have not hitherto been addressable. Finally, the precision afforded by the automated robotic control system will allow the patch-clamping of subcellular structures in vivo, which has not previously been systematically achievable. We will demonstrate the utility of our two-photon targeted robotic patch clamp platform by using it to target two particular classes of pyramidal cell in the mouse cerebral cortex, in order to ascertain their respective roles in processing sensory information.

全细胞膜片钳记录技术通过记录离子通道中的电流在生命科学中产生了很大的影响：这对我们理解细胞功能的进步至关重要。在体内使用膜片钳技术，然而，一直很困难，因为它基本上是进行“盲”，与唯一的反馈，通过监测用于使recording.Recent发展中的多光子显微镜的微管阻抗获得现在允许我们从已被荧光标记的细胞进行有针对性的记录。最近还表明，在体内膜片钳记录可以自动化。在本计画中，我们将联合收割机结合这些原理，发展出一种全细胞膜片钳电生理学的新方法。我们的平台将允许人类操作员视觉选择（“点击”）遗传靶向细胞类别，然后由一组能够同时获得多达六个细胞记录的机器人显微操作器自动修补。除了允许用于基础科学或药物发现目的的体内细胞的高通量表征外，我们的平台还将允许提出新的科学问题，涉及迄今为止尚未解决的细胞之间的相互作用。最后，自动化机器人控制系统提供的精度将允许在体内对亚细胞结构进行膜片钳，这在以前是无法系统实现的。我们将展示我们的双光子靶向机器人膜片钳平台的实用性，通过使用它来靶向小鼠大脑皮层中的两个特定类别的锥体细胞，以确定它们各自在处理感觉信息中的作用。

项目成果

NeuroFlow: A General Purpose Spiking Neural Network Simulation Platform using Customizable Processors.

• DOI: 10.3389/fnins.2015.00516
• 发表时间: 2015
• 期刊: Frontiers in neuroscience
• 影响因子: 4.3
• 作者: Cheung K;Schultz SR;Luk W
• 通讯作者: Luk W

Tectal-derived interneurons contribute to phasic and tonic inhibition in the visual thalamus.

• DOI: 10.1038/ncomms13579
• 发表时间: 2016-12-08
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Jager, Polona;Ye, Zhiwen;Yu, Xiao;Zagoraiou, Laskaro;Prekop, Hong-Ting;Partanen, Juha;Jessell, Thomas M.;Wisden, William;Brickley, Stephen G.;Delogu, Alessio
• 通讯作者: Delogu, Alessio

Two-photon targeted robotic patch-clamp electrophysiological recording in vivo.

双光子靶向机器人膜片钳体内电生理记录。

• 发表时间: 2013
• 期刊: Society for Neuroscience. Online
• 作者: Annecchino, LA

Progress in automating patch clamp cellular physiology.

• DOI: 10.1177/2398212818776561
• 发表时间: 2018-01-01
• 期刊: Brain and neuroscience advances
• 作者: Annecchino, Luca A;Schultz, Simon R
• 通讯作者: Schultz, Simon R

Neuronal gain modulability is determined by dendritic morphology: a computational optogenetic study

神经元增益可调节性由树突形态决定：计算光遗传学研究

• DOI: 10.1101/096586
• 发表时间: 2016
• 作者: Jarvis S