Garaschuk Olga, Milos Ruxandra-Iulia, Grienberger Christine, Marandi Nima, Adelsberger Helmuth, Konnerth Arthur
Institut für Neurowissenschaften, Technische Universität München, Biedersteinerstr. 29, 80802, Munich, Germany.
Pflugers Arch. 2006 Dec;453(3):385-96. doi: 10.1007/s00424-006-0150-x. Epub 2006 Oct 18.
The precise understanding of the cellular and molecular basis of brain function requires the direct assessment of the activity of defined cells in vivo. A promising approach for such analyses is two-photon microscopy in combination with appropriate cell labeling techniques. Here, we review the multi-cell bolus loading (MCBL) method that involves the use of membrane-permeant fluorescent indicator dyes. We show that this approach is useful for the functional analysis of clusters of neurons and glial cells in vivo. Work from our and other laboratories shows that the techniques that were previously feasible only in brain slices, like targeted patch clamp recordings from identified cells or pharmacological manipulations in confined brain regions, can now be used also in vivo. We also show that MCBL and two-photon imaging can be easily combined with other labeling techniques, particularly with those involving the use of genetically encoded, green-fluorescent-protein-based indicators. Finally, we examine recent applications of MCBL/two-photon imaging for the analysis of various brain regions, including the somatosensory and the visual cortex.
要精确理解脑功能的细胞和分子基础，需要在体内直接评估特定细胞的活性。一种很有前景的分析方法是将双光子显微镜与适当的细胞标记技术相结合。在此，我们回顾了涉及使用膜通透性荧光指示剂染料的多细胞团块加载（MCBL）方法。我们表明，这种方法对于体内神经元和神经胶质细胞簇的功能分析很有用。我们实验室和其他实验室的研究表明，以前仅在脑切片中可行的技术，如从特定细胞进行靶向膜片钳记录或在有限脑区进行药理学操作，现在也可用于体内。我们还表明，MCBL和双光子成像可以很容易地与其他标记技术相结合，特别是与那些涉及使用基于绿色荧光蛋白的基因编码指示剂的技术。最后，我们研究了MCBL/双光子成像在分析包括体感皮层和视觉皮层在内的各种脑区方面的最新应用。
