Adaptive optics enables multimode 3D super-resolution microscopy via remote focusing
Vytautas Navikas (École Polytechnique Fédérale de Lausanne)
Adrien C. Descloux (École Polytechnique Fédérale de Lausanne)
Kristin Grusmayer (Kavli institute of nanoscience Delft, TU Delft - BN/Kristin Grussmayer Lab, École Polytechnique Fédérale de Lausanne)
Sanjin Marion (École Polytechnique Fédérale de Lausanne)
Aleksandra Radenovic (École Polytechnique Fédérale de Lausanne)
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Abstract
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.
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