美国加州大学圣巴巴拉分校Spencer LaVere Smith和Che-Hang Yu共同合作，近期取得重要工作进展。他们研究开发了Cousa物镜，这是一种用于多光子体内成像的长工作距离空气物镜。相关研究成果2023年12月21日在线发表于《自然—方法学》杂志上。

据介绍，多光子显微镜可以解析散射组织深处的荧光结构和动力学，并改变了神经成像，但在体内应用这项技术可能会受到传统物镜的机械和光学限制。短工作距离物镜可能会与紧凑的手术窗或其他仪器发生碰撞，从而妨碍成像。

研究人员开发了超长工作距离（20 mm）空气物镜，称为Cousa物镜。它针对多光子成像波长的性能进行了优化，提供了超过4 mm²视场，具有亚微米横向分辨率，并与常用的多光子成像系统兼容。一种新颖的机械设计，比典型的显微镜物镜更宽，使这种规格组合成为可能。研究人员分享了完整的光学处方，并报告了包括非人类灵长类动物在内的一系列物种和制剂的体内双光子和三光子成像在内的性能。

总之，Cousa物镜可以实现神经科学及其它领域的一系列实验。

附：英文原文

Title: The Cousa objective: a long-working distance air objective for multiphoton imaging in vivo

Author: Yu, Che-Hang, Yu, Yiyi, Adsit, Liam M., Chang, Jeremy T., Barchini, Jad, Moberly, Andrew H., Benisty, Hadas, Kim, Jinkyung, Young, Brent K., Heng, Kathleen, Farinella, Deano M., Leikvoll, Austin, Pavan, Rishaab, Vistein, Rachel, Nanfito, Brandon R., Hildebrand, David G. C., Otero-Coronel, Santiago, Vaziri, Alipasha, Goldberg, Jeffrey L., Ricci, Anthony J., Fitzpatrick, David, Cardin, Jessica A., Higley, Michael J., Smith, Gordon B., Kara, Prakash, Nielsen, Kristina J., Smith, Ikuko T., Smith, Spencer LaVere

Issue&Volume: 2023-12-21

Abstract: Multiphoton microscopy can resolve fluorescent structures and dynamics deep in scattering tissue and has transformed neural imaging, but applying this technique in vivo can be limited by the mechanical and optical constraints of conventional objectives. Short working distance objectives can collide with compact surgical windows or other instrumentation and preclude imaging. Here we present an ultra-long working distance (20mm) air objective called the Cousa objective. It is optimized for performance across multiphoton imaging wavelengths, offers a more than 4mm2 field of view with submicrometer lateral resolution and is compatible with commonly used multiphoton imaging systems. A novel mechanical design, wider than typical microscope objectives, enabled this combination of specifications. We share the full optical prescription, and report performance including in vivo two-photon and three-photon imaging in an array of species and preparations, including nonhuman primates. The Cousa objective can enable a range of experiments in neuroscience and beyond.

DOI: 10.1038/s41592-023-02098-1

Source: https://www.nature.com/articles/s41592-023-02098-1