Aberrations and adaptive optics in super-resolution microscopy

AbstractMicroscope-AOtools is a software package which allows for a simple, robust and generalised implementation of adaptive optics (AO) elements. It contains all the necessary methods for set-up, calibration, and aberration correction which are simple to use and function in a robust manner. Aberrations arising from sources such as sample hetero-geneity and refractive index mismatches are constant problems in biological imaging. These aberrations reduce image quality and the achievable depth of imaging, particularly in super-resolution microscopy techniques. AO technology has been proven to be effective in correcting for these aberrations and thereby improving the image quality. However, it has not been widely adopted by the biological imaging community due, in part, to difficulty in set-up and operation of AO, particularly by non-specialist users. Microscope-AOtools offers a robust, easy-to-use implementation of the essential methods for set-up and use of AO techniques. These methods are constructed in a generalised manner that can utilise a range of adaptive optics elements, wavefront sensing techniques and sensorless AO correction methods. Furthermore, the methods are designed to be easily extensible as new techniques arise, leading to a streamlined pipeline for new AO technology and techniques to be adopted by the wider microscopy community.

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Active PSF Shaping and Adaptive Optics Enable Volumetric Single Molecule Super-Resolution Microscopy through Brain Sections

Biophysical Journal ◽

10.1016/j.bpj.2018.11.1533 ◽

2019 ◽

Vol 116 (3) ◽

pp. 283a-284a

Author(s):

Michael Mlodzianoski ◽

Paul Cheng-Hathaway ◽

Sheng Liu ◽

Shane Bemiller ◽

Tyler McCray ◽

...

Keyword(s):

Adaptive Optics ◽

Single Molecule ◽

Super Resolution ◽

Super Resolution Microscopy

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Dynamic super-resolution structured illumination imaging in the living brain

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1819965116 ◽

2019 ◽

Vol 116 (19) ◽

pp. 9586-9591 ◽

Cited By ~ 21

Author(s):

Raphaël Turcotte ◽

Yajie Liang ◽

Masashi Tanimoto ◽

Qinrong Zhang ◽

Ziwei Li ◽

...

Keyword(s):

Adaptive Optics ◽

Super Resolution ◽

Structured Illumination ◽

Structured Illumination Microscopy ◽

Optical Aberrations ◽

Sample Motion ◽

Heterogeneous Tissue ◽

Conventional Microscopy ◽

Super Resolution Microscopy

Cells in the brain act as components of extended networks. Therefore, to understand neurobiological processes in a physiological context, it is essential to study them in vivo. Super-resolution microscopy has spatial resolution beyond the diffraction limit, thus promising to provide structural and functional insights that are not accessible with conventional microscopy. However, to apply it to in vivo brain imaging, we must address the challenges of 3D imaging in an optically heterogeneous tissue that is constantly in motion. We optimized image acquisition and reconstruction to combat sample motion and applied adaptive optics to correcting sample-induced optical aberrations in super-resolution structured illumination microscopy (SIM) in vivo. We imaged the brains of live zebrafish larvae and mice and observed the dynamics of dendrites and dendritic spines at nanoscale resolution.

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Optimal Sensorless Adaptive Optics Schemes for Super-Resolution Microscopy

Imaging and Applied Optics ◽

10.1364/aopt.2013.otu1a.2 ◽

2013 ◽

Author(s):

Daniel Burke ◽

Fiona Kenny ◽

Brian Patton ◽

Martin J. Booth

Keyword(s):

Adaptive Optics ◽

Super Resolution ◽

Super Resolution Microscopy

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

Nanophotonics ◽

10.1515/nanoph-2021-0108 ◽

2021 ◽

Vol 0 (0) ◽

Author(s):

Vytautas Navikas ◽

Adrien C. Descloux ◽

Kristin S. Grussmayer ◽

Sanjin Marion ◽

Aleksandra Radenovic

Keyword(s):

Adaptive Optics ◽

Single Molecule ◽

Biological Sample ◽

Super Resolution ◽

Axial Position ◽

Deformable Mirror ◽

Wavefront Control ◽

Convenient Approach ◽

Wavefront Shaping ◽

Super Resolution Microscopy

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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Super-Resolution Microscopy in Studying the Structure and Function of the Cell Nucleus

Acta Naturae ◽

10.32607/2075851-2017-9-4-42-51 ◽

2017 ◽

Vol 9 (4) ◽

pp. 42-51

Author(s):

S. S. Ryabichko ◽

◽

A. N. Ibragimov ◽

L. A. Lebedeva ◽

E. N. Kozlov ◽

...

Keyword(s):

Cell Nucleus ◽

Super Resolution ◽

Structure And Function ◽

And Function ◽

Super Resolution Microscopy

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Structural Evidence of Photoisomerization Pathways in Fluorescent Proteins

10.26434/chemrxiv.9209450.v1 ◽

2019 ◽

Author(s):

Jeffrey Chang ◽

Matthew Romei ◽

Steven Boxer

Keyword(s):

Rational Design ◽

Fluorescent Protein ◽

Fluorescent Proteins ◽

Super Resolution ◽

Unit Cell Size ◽

Chlorine Substituent ◽

Gfp Chromophore ◽

The One ◽

Green Fluorescent ◽

Super Resolution Microscopy

Double-bond photoisomerization in molecules such as the green fluorescent protein (GFP) chromophore can occur either via a volume-demanding one-bond-flip pathway or via a volume-conserving hula-twist pathway. Understanding the factors that determine the pathway of photoisomerization would inform the rational design of photoswitchable GFPs as improved tools for super-resolution microscopy. In this communication, we reveal the photoisomerization pathway of a photoswitchable GFP, rsEGFP2, by solving crystal structures of cis and trans rsEGFP2 containing a monochlorinated chromophore. The position of the chlorine substituent in the trans state breaks the symmetry of the phenolate ring of the chromophore and allows us to distinguish the two pathways. Surprisingly, we find that the pathway depends on the arrangement of protein monomers within the crystal lattice: in a looser packing, the one-bond-flip occurs, whereas in a tighter packing (7% smaller unit cell size), the hula-twist occurs.

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