Wide-field and two-photon imaging of brain activity with voltage- and calcium-sensitive dyes
Wide-field and two-photon imaging of brain activity with voltage- and calcium-sensitive dyes
复制标题
DOI：
10.1098/rstb.2009.0084
复制
发表时间：
2009-09-12
影响因子：
6.3
通讯作者：
Zecevic, Dejan
中科院分区：
文献类型：
--
作者：
Homma, Ryota;Baker, Bradley J.;Zecevic, Dejan
这篇综述介绍了三个使用电压或钙敏感染料来成像大脑活动的例子。我们的目的是讨论每种方法的优点和缺点，特别是其应用于脑干的研究。其中两个例子使用宽场（单光子）成像;第三个使用双光子扫描显微镜。由于测量具有有限的信噪比，本文还讨论了优化信号的关键方法。以下三个例子。(i)细胞内注射的电压敏感染料被用来监测膜电位在体外制备的神经元树突。这些实验旨在了解单个神经元如何将复杂的突触输入转换为输出尖峰序列。(ii)细胞外，浴应用电压敏感染料被用来监测人口信号从不同部分的背侧脑干。我们描述了呼吸活动期间的记录。群体信号表明四个不同的区域具有与吸气相关的不同活动。(iii)钙敏感染料可用于标记哺乳动物大脑中的许多单个细胞。这种方法与双光子显微镜相结合，使得在虚构的呼吸节律过程中跟踪体外脑干标本中的尖峰活动成为可能。今天使用的有机电压和离子敏感染料不加区别地染色标本中的所有细胞类型。一个主要的努力正在进行中，以开发荧光蛋白传感器的活性选择性染色个别细胞类型。
This review presents three examples of using voltage- or calcium-sensitive dyes to image the activity of the brain. Our aim is to discuss the advantages and disadvantages of each method with particular reference to its application to the study of the brainstem. Two of the examples use wide-field (one-photon) imaging; the third uses two-photon scanning microscopy. Because the measurements have limited signal-to-noise ratio, the paper also discusses the methodological aspects that are critical for optimizing the signal.The three examples are the following. (i) An intracellularly injected voltage-sensitive dye was used to monitor membrane potential in the dendrites of neurons in in vitro preparations. These experiments were directed at understanding how individual neurons convert complex synaptic inputs into the output spike train. (ii) An extracellular, bath application of a voltage-sensitive dye was used to monitor population signals from different parts of the dorsal brainstem. We describe recordings made during respiratory activity. The population signals indicated four different regions with distinct activity correlated with inspiration. (iii) Calcium-sensitive dyes can be used to label many individual cells in the mammalian brain. This approach, combined with two-photon microscopy, made it possible to follow the spike activity in an in vitro brainstem preparation during fictive respiratory rhythms.The organic voltage-and ion-sensitive dyes used today indiscriminatively stain all of the cell types in the preparation. A major effort is underway to develop fluorescent protein sensors of activity for selectively staining individual cell types.