...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >In vivo two-photon calcium imaging of neuronal networks
【24h】

In vivo two-photon calcium imaging of neuronal networks

机译：神经元网络的体内双光子钙成像

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Two-photon calcium imaging is a powerful means for monitoring the activity of distinct neurons in brain tissue in vivo. In the mammalian brain, such imaging studies have been restricted largely to calcium recordings from neurons that were individually dye-loaded through microelectrodes. Previous attempts to use membrane-permeant forms of fluorometric calcium indicators to load populations of neurons have yielded satisfactory results only in cell cultures or in slices of immature brain tissue. Here we introduce a versatile approach for loading membrane-permeant fluorescent indicator dyes in large populations of cells. We established a pressure ejection-based local dye delivery protocol that can be used for a large spectrum of membrane-permeant indicator dyes, including calcium green-1 acetoxymethyl (AM) ester, Fura-2 AM, Fluo-4 AM, and Indo-1 AM. We applied this dye-loading protocol successfully in mouse brain tissue at any developmental stage from newborn to adult in vivo and in vitro. In vivo two-photon Ca2+ recordings, obtained by imaging through the intact skull, indicated that whisker deflection-evoked Ca2+ transients occur in a subset of layer 2/3 neurons of the barrel cortex. Thus, our results demonstrate the suitability of this technique for real-time analyses of intact neuronal circuits with the resolution of individual cells. [References: 42]
机译：两光子钙成像是监测体内脑组织中不同神经元活动的有力手段。在哺乳动物的大脑中，此类成像研究在很大程度上局限于来自神经元的钙记录，而这些神经元是通过微电极单独上染的。以前尝试使用透过膜形式的荧光钙指示剂来加载神经元种群的尝试仅在细胞培养物中或未成熟的脑组织切片中产生了令人满意的结果。在这里，我们介绍了一种用于在大量细胞中加载膜渗透性荧光指示剂染料的通用方法。我们建立了基于压力喷射的局部染料传输协议，该协议可用于大范围的膜渗透指示剂染料，包括绿钙1乙酰氧基甲基（AM）酯，Fura-2 AM，Fluo-4 AM和Indo-凌晨1点。我们在体内和体外从新生到成年的任何发育阶段都成功地将这种染料加载方案应用于小鼠脑组织。通过完整的颅骨成像获得的体内双光子Ca2 +记录表明，晶须偏转诱发的Ca2 +瞬变发生在桶状皮质的第2/3层神经元的子集中。因此，我们的结果证明了该技术适用于具有单个细胞分辨率的完整神经回路实时分析。 [参考：42]

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号