Adaptive optics enables multimode 3D super-resolution microscopy via remote focusing

Nanophotonics

article

Vytautas Navikas¹  Adrien C. Descloux¹  Kristin S. Grussmayer¹  Sanjin Marion¹  Aleksandra Radenovic¹ 

[1] Laboratory of Nanoscale Biology, Institute of Bioengineering, School of Engineering, Swiss Federal Institute of Technology Lausanne (EPFL);Grussmayer Lab, Department of Bionanoscience, Faculty of Applied Science, Kavli Institute for Nanoscience Delft, Delft University of Technology

关键词: adaptive optics;    astigmatism-based single molecule localization microscopy;    3D imaging;    remote focusing;    single-molecule localization microscopy (SMLM);    super-resolution optical fluctuation imaging (SOFI);   

DOI  :  10.1515/nanoph-2021-0108

学科分类：社会科学、人文和艺术（综合）

来源: De Gruyter

【 摘 要 】

A variety of modern super-resolution microscopy methods provide researchers with previously inconceivable biological sample imaging opportunities at a molecular resolution. All of these techniques excel at imaging samples that are close to the coverslip, however imaging at large depths remains a challenge due to aberrations caused by the sample, diminishing the resolution of the microscope. Originating in astro-imaging, the adaptive optics (AO) approach for wavefront shaping using a deformable mirror is gaining momentum in modern microscopy as a convenient approach for wavefront control. AO has the ability not only to correct aberrations but also enables engineering of the PSF shape, allowing localization of the emitter axial position over several microns. In this study, we demonstrate remote focusing as another AO benefit for super-resolution microscopy. We show the ability to record volumetric data (45 × 45 × 10 µm), while keeping the sample axially stabilized using a standard widefield setup with an adaptive optics addon. We processed the data with single-molecule localization routines and/or computed spatiotemporal correlations, demonstrating subdiffraction resolution.

【 授权许可】

CC BY