Quirin Sean
Opt Express. 2025 Jan 27;33(2):1647-1659. doi: 10.1364/OE.544906.
Biological applications using multiphoton microscopy increasingly seek a larger field of view while maintaining sufficient temporal sampling to observe dynamic biological processes. Multiphoton imaging also requires high numerical aperture microscope objectives to realize efficient non-linear excitation and collection of fluorescence. This combination of low-magnification and high-numerical aperture poses a challenge for system design. To address this, the use of a liquid crystal polarization grating stack is proposed here to temporally sequence through multiple fields of view. This solution pans the native field of view with minimal latency and zero inertial movement of either the microscope or biological sample. Implemented as a simple add-on unit to existing multi-photon microscopes, this device increases the total field size by 4x, covering up to 7.6mm. Performance constraints and functional demonstration of imaging neural activity are presented.
使用多光子显微镜的生物学应用越来越多地寻求更大的视野，同时保持足够的时间采样以观察动态生物学过程。多光子成像还需要高数值孔径的显微镜物镜来实现高效的非线性激发和荧光收集。这种低放大倍数和高数值孔径的组合给系统设计带来了挑战。为了解决这个问题，本文提出使用液晶偏振光栅堆栈对多个视野进行时间排序。该解决方案以最小的延迟和平移显微镜或生物样品的零惯性运动来扫视原生视野。作为现有多光子显微镜的一个简单附加单元实现，该设备将总视野大小增加了4倍，覆盖范围可达7.6毫米。本文还介绍了成像神经活动的性能限制和功能演示。
