Fast 3D multiphoton microscope

该应用程序是一个超快多光子成像平台，融合了三维（3D）荧光采集和（目标）三维光刺激。我们研究鼠脑或人类诱导多能干细胞（iPSCs）衍生的脑类器官，以了解神经退行性疾病的基本机制，这些器官包含通过精细过程连接的神经元和胶质细胞的3D网络。可视化这些复杂的结构，特别是在3D中检测同时发生在分散位置的快速时间事件（例如，穿过神经元树突树的棘）需要非常快速的扫描。这可以通过使用声光（AO）技术利用千赫采样率，允许随机接入点扫描。结合荧光钙指示剂，可以在大体积内研究数百个相互连接的神经元的活动，包括相对稀疏的神经元群（如抑制性中间神经元）的成像。这对于同时分析大量不同神经元亚型的活动变化的后果或细胞亚群中细胞内蛋白质聚集的后果尤为重要。由于扫描速率的限制，传统的基于galvo /谐振的多光子逐平面成像无法实现这一点。

This application is for an ultrafast multiphoton imaging platform that fuses three-dimensional (3D) fluorescence acquisition with (targeted) 3D photostimulation. The mouse brain or human induced pluripotent stem cells (iPSCs) derived brain organoids which we study to understand basic mechanisms of neurodegenerative diseases contain 3D networks of neurons and glia cells connected via fine processes. Visualizing such elaborate structures in action, in particular detecting fast temporal events occurring simultaneously at dispersed locations in 3D (e.g., across spines on a neuronal dendritic tree) need very fast scanning. This can be utilized by the use of acousto-optic (AO) technology with kHz sampling rates which allow random-access point scanning. In combination with fluorescent calcium indicators, the activity of hundreds of interconnected neurons can be studied within large volumes including the imaging of relatively sparse neuronal populations such as inhibitory interneurons. This is especially important in order to simultaneously analyse the consequence of activity changes in a multitude of different neuronal subtypes or the consequence of intracellular protein aggregates in a subset of cells. Due to its scanning rate limitations, this cannot be achieved with conventional galvo /resonant based multiphoton plane-by-plane imaging.