Online correction of licking-induced brain motion during two-photon imaging with a tunable lens
Online correction of licking-induced brain motion during two-photon imaging with a tunable lens
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DOI：
10.1113/jphysiol.2013.259804
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发表时间：
2013-10-01
影响因子：
5.5
通讯作者：
Helmchen, Fritjof
中科院分区：
文献类型：
--
作者：
Chen, Jerry L.;Pfaeffli, Oliver A.;Helmchen, Fritjof
在清醒、头部固定的动物中进行双光子钙成像，可以测量行为期间的神经元活动。通常情况下，舔水奖励检索被用作动物的决定在奖励驱动的行为可衡量的报告。然而，舔行为会引起严重的运动伪影，干扰细胞活动的双光子成像。在这里，我们描述了一个简单的方法，在线校正舔引起的焦点转移的双光子钙成像的新皮层神经元在头部固定的小鼠。我们发现，舔导致新皮层组织的刻板下降，将神经元转移到20米以外的焦点。基于压电薄膜传感器的舔测量，我们开发了一个反馈模型，它提供了一个校正信号的快速光学聚焦调整与电可调透镜。使用在线校正与此反馈模型，我们证明了减少舔相关的焦点变化低于3米，最大限度地减少运动伪影污染的细胞钙信号。利用可调透镜进行聚焦校正是一种简单有效的方法，可以提高在基于奖励的行为期间监测神经元活动的能力。
Two-photon calcium imaging in awake, head-fixed animals enables the measurement of neuronal activity during behaviour. Often, licking for the retrieval of water reward is used as a measurable report of the animal's decision during reward-driven behaviour. However, licking behaviour can induce severe motion artifacts that interfere with two-photon imaging of cellular activity. Here, we describe a simple method for the online correction of licking-induced focus shifts for two-photon calcium imaging of neocortical neurons in the head-fixed mouse. We found that licking causes a stereotyped drop of neocortical tissue, shifting neurons up to 20 m out of focus. Based on the measurement of licking with a piezo film sensor, we developed a feedback model, which provides a corrective signal for fast optical focus adjustments with an electrically tunable lens. Using online correction with this feedback model, we demonstrate a reduction of licking-related focus changes below 3 m, minimizing motion artifact contamination of cellular calcium signals. Focus correction with a tunable lens is a simple and effective method to improve the ability to monitor neuronal activity during reward-based behaviour.