Institute of Experimental Medicine, Hungarian Academy of Sciences, Budapest, Hungary.
Nat Methods. 2012 Jan 8;9(2):201-8. doi: 10.1038/nmeth.1851.
The understanding of brain computations requires methods that read out neural activity on different spatial and temporal scales. Following signal propagation and integration across a neuron and recording the concerted activity of hundreds of neurons pose distinct challenges, and the design of imaging systems has been mostly focused on tackling one of the two operations. We developed a high-resolution, acousto-optic two-photon microscope with continuous three-dimensional (3D) trajectory and random-access scanning modes that reaches near-cubic-millimeter scan range and can be adapted to imaging different spatial scales. We performed 3D calcium imaging of action potential backpropagation and dendritic spike forward propagation at sub-millisecond temporal resolution in mouse brain slices. We also performed volumetric random-access scanning calcium imaging of spontaneous and visual stimulation-evoked activity in hundreds of neurons of the mouse visual cortex in vivo. These experiments demonstrate the subcellular and network-scale imaging capabilities of our system.
理解大脑计算需要能够在不同时空尺度上读取神经活动的方法。沿着神经元进行信号传播和整合，并记录数百个神经元的协同活动，这带来了明显的挑战，因此成像系统的设计主要集中在解决这两个操作中的一个。我们开发了一种高分辨率、声光双光子显微镜，具有连续的三维（3D）轨迹和随机访问扫描模式，可达到近立方毫米的扫描范围，并可适应不同空间尺度的成像。我们在小鼠脑片上以亚毫秒时间分辨率进行了动作电位逆行传播和树突棘前向传播的 3D 钙成像。我们还在活体小鼠视觉皮层的数百个神经元中进行了自发和视觉刺激诱发活动的容积随机访问扫描钙成像。这些实验证明了我们系统的亚细胞和网络尺度成像能力。
