Heuristically optimal path scanning for high-speed multiphoton circuit imaging
Heuristically optimal path scanning for high-speed multiphoton circuit imaging
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DOI：
10.1152/jn.00334.2011
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发表时间：
2011-09-01
影响因子：
2.5
通讯作者：
MacLean, Jason N.
中科院分区：
文献类型：
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作者：
Sadovsky, Alexander J.;Kruskal, Peter B.;MacLean, Jason N.
Sadovsky AJ，Kruskal PB，Kimmel JM，Ostmeyer J，Neubauer FB，MacLean JN。用于高速多光子电路成像的启发式最佳路径扫描。神经生理学杂志106：1591-1598,2011。2011年6月29日首次出版；doi：10.1152/jn.00334.2011。-有模式的神经元放电的群体动力学是大脑信息处理的基础。多光子显微镜与钙指示剂染料相结合，可以用单神经元分辨率对电路动态进行成像。然而，荧光测量的时间分辨率受到标准栅格扫描技术施加的成像频率的限制。因此，传统的栅格扫描限制了在成像的神经元群体中检测动作电位的相对时序的能力。为了使用标准的多光子激光扫描显微镜(MPLSM)装置最大限度地提高来自大量神经元的荧光测量速度，我们开发了启发式最优路径扫描(HOPS)。使用标准检流计扫描镜，HOPS优化了激光传播路径长度，从而提高了神经元荧光测量的时间分辨率。仅将扫描路径最小化不足以对神经元群进行长时间的高速成像。随着扫描速率的增加，路径稳定性和信噪比变得越来越重要。HOPS通过表征扫描镜振镜来解决这一问题，以实现更长的路径稳定性。此外，优化了神经元驻留时间，在最大化扫描速度的同时提高了对动作电位的检测能力。最短路径计算和最小化镜像定位时间的组合使我们能够以单脉冲分辨率以高速率光学监控视野中的神经元群体，类似于50个神经元的125赫兹和类似于1,000个神经元的8.5赫兹。我们的方法引入了一种可访问的方法，使用传统的MPLSM对大量神经元群体进行快速成像，有助于对神经元电路动力学的新见解。
Sadovsky AJ, Kruskal PB, Kimmel JM, Ostmeyer J, Neubauer FB, MacLean JN. Heuristically optimal path scanning for high-speed multiphoton circuit imaging. J Neurophysiol 106: 1591-1598, 2011. First published June 29, 2011; doi: 10.1152/jn.00334.2011.-Population dynamics of patterned neuronal firing are fundamental to information processing in the brain. Multiphoton microscopy in combination with calcium indicator dyes allows circuit dynamics to be imaged with single-neuron resolution. However, the temporal resolution of fluorescent measures is constrained by the imaging frequency imposed by standard raster scanning techniques. As a result, traditional raster scans limit the ability to detect the relative timing of action potentials in the imaged neuronal population. To maximize the speed of fluorescence measures from large populations of neurons using a standard multiphoton laser scanning microscope (MPLSM) setup, we have developed heuristically optimal path scanning (HOPS). HOPS optimizes the laser travel path length, and thus the temporal resolution of neuronal fluorescent measures, using standard galvanometer scan mirrors. Minimizing the scan path alone is insufficient for prolonged high-speed imaging of neuronal populations. Path stability and the signal-to-noise ratio become increasingly important factors as scan rates increase. HOPS addresses this by characterizing the scan mirror galvanometers to achieve prolonged path stability. In addition, the neuronal dwell time is optimized to sharpen the detection of action potentials while maximizing scan rate. The combination of shortest path calculation and minimization of mirror positioning time allows us to optically monitor a population of neurons in a field of view at high rates with single-spike resolution, similar to 125 Hz for 50 neurons and similar to 8.5 Hz for 1,000 neurons. Our approach introduces an accessible method for rapid imaging of large neuronal populations using traditional MPLSMs, facilitating new insights into neuronal circuit dynamics.