美国耶鲁大学Joerg Bewersdorf课题组开发出可用于厚样本成像的三维自适应光学纳米显微镜。2021年5月31日，《自然—方法学》杂志在线发表了这项成果。

研究人员开发了一种改进的等激发发射耗尽纳米显微镜，其使用先进的自适应光学策略来实现低于50nm分辨率，例如在组织中以前无法到达的神经元突触和环形管。该自适应光学方案通常适用于具有类似光束路径几何形状的任何显微镜，即涉及两个相对的物镜，从而可在深处成像时优化分辨率。

据悉，了解细胞结构需要所有三个维度的最佳空间分辨率。在荧光显微镜中，这是通过4Pi纳米镜方法实现的，该方法将两个相对物镜用于最佳衍射限制3D分辨率 ，并在明亮和暗态之间切换荧光分子来破坏衍射极限。然而，光学像差将这些纳米镜限制于薄样品并阻止了其在厚样本中的应用。

附：英文原文

Title: Three-dimensional adaptive optical nanoscopy for thick specimen imaging at sub-50-nm resolution

Author: Xiang Hao, Edward S. Allgeyer, Dong-Ryoung Lee, Jacopo Antonello, Katherine Watters, Julianne A. Gerdes, Lena K. Schroeder, Francesca Bottanelli, Jiaxi Zhao, Phylicia Kidd, Mark D. Lessard, James E. Rothman, Lynn Cooley, Thomas Biederer, Martin J. Booth, Joerg Bewersdorf

Issue&Volume: 2021-05-31

Abstract: Understanding cellular organization demands the best possible spatial resolution in all three dimensions. In fluorescence microscopy, this is achieved by 4Pi nanoscopy methods that combine the concepts of using two opposing objectives for optimal diffraction-limited 3D resolution with switching fluorescent molecules between bright and dark states to break the diffraction limit. However, optical aberrations have limited these nanoscopes to thin samples and prevented their application in thick specimens. Here we have developed an improved iso-stimulated emission depletion nanoscope, which uses an advanced adaptive optics strategy to achieve sub-50-nm isotropic resolution of structures such as neuronal synapses and ring canals previously inaccessible in tissue. The adaptive optics scheme presented in this work is generally applicable to any microscope with a similar beam path geometry involving two opposing objectives to optimize resolution when imaging deep in aberrating specimens. The combination of adaptive optics with an improved isoSTED nanoscope allows imaging of cells and tissues with sub-50-nm isotropic resolution.

DOI: 10.1038/s41592-021-01149-9

Source: https://www.nature.com/articles/s41592-021-01149-9