Precision Calcium Imaging of Dense Neural Populations via a Cell-Body-Targeted Calcium Indicator
Precision Calcium Imaging of Dense Neural Populations via a Cell-Body-Targeted Calcium Indicator
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DOI：
10.1016/j.neuron.2020.05.029
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发表时间：
2020-08-05
期刊：
影响因子：
16.2
通讯作者：
Boyden, Edward S.
中科院分区：
文献类型：
--
作者：
Shemesh, Or A.;Linghu, Changyang;Boyden, Edward S.
钙动力学的单光子荧光成像方法可以以较低的设备复杂性和成本捕获大视野中数百个神经元的活动。然而，与双光子方法相比，单光子方法受到更高水平的神经脉冲串扰的影响，导致信噪比降低，神经活动的人工相关性降低。我们通过设计针对细胞体的荧光钙指示剂GCaMP6f和GCaMP7f的变体来解决这个问题。我们筛选了GCaMP与天然和人造多肽的融合，并鉴定了将GCaMP定位在小鼠和斑马鱼幼体神经元细胞体50 mm以内的融合。在密集的神经回路中，以Soma为靶点的GCaMP的单光子成像报告了来自神经纤维的伪性尖峰减少，信噪比增加，以及神经元之间的伪相关性降低。因此，以SOMA为靶点的荧光钙指示剂有助于使用简单、强大的单光子方法来成像神经钙动力学。
Methods for one-photon fluorescent imaging of calciumdynamics can capture the activity of hundreds of neurons across large fields of view at a low equipment complexity and cost. In contrast to two-photon methods, however, one-photon methods suffer from higher levels of crosstalk from neuropil, resulting in a decreased signal-to-noise ratio and artifactual correlations of neural activity. We address this problem by engineering cell-body-targeted variants of the fluorescent calcium indicators GCaMP6f and GCaMP7f. We screened fusions of GCaMP to natural, as well as artificial, peptides and identified fusions that localized GCaMP to within 50 mmof the cell body of neurons in mice and larval zebrafish. One-photon imaging of soma-targetedGCaMPin dense neural circuits reported fewer artifactual spikes from neuropil, an increased signal-to-noise ratio, and decreased artifactual correlation across neurons. Thus, soma-targeting of fluorescent calcium indicators facilitates usage of simple, powerful, one-photon methods for imaging neural calcium dynamics.