scholarly journals Numerically enhanced adaptive optics-based 3D STED microscopy for deep-tissue super-resolved imaging

2019 ◽  
Author(s):  
Piotr Zdankowski ◽  
Maciej Trusiak ◽  
David McGloin ◽  
Jason R. Swedlow
Keyword(s):  
Adaptive Optics ◽  
Signal To Noise Ratio ◽  
Super Resolution ◽  
Thick Layer ◽  
Block Matching ◽  
Stimulated Emission ◽  
Optical Aberrations ◽  
Deep Tissue ◽  
Resolution Imaging ◽  
Induced Pluripotent

AbstractIn stimulated emission depletion (STED) nanoscopy, the major origin of decreased signal-to-noise ratio within images can be attributed to sample photobleaching and strong optical aberrations. This is due to STED utilising both a high power depletion laser (increasing risk of photodamage), while the depletion beam is very sensitive to sample-induced aberrations. Here we demonstrate a custom-built 3D STED microscope with automated aberration correction that is capable of 3D super-resolution imaging through thick, highly aberrating, tissue. We introduce and investigate image denoising by block-matching and collaborative filtering (BM3D) to numerically enhance fine object details otherwise mixed with noise. Numerical denoising provides an increase in the final effective resolution of the STED imaging of 31% using the well-established Fourier ring correlation metric. Experimental validation of the proposed method is achieved through super-resolved 3D imaging of axons in differentiated induced pluripotent stem cells growing under a 80µm thick layer of tissue with lateral and axial resolution of 256nm and 300nm, respectively.

scholarly journals Adaptive optics allows 3D STED-FCS measurements in the cytoplasm of living cells

2019 ◽  
Author(s):  
Aurélien Barbotin ◽  
Silvia Galiani ◽  
Iztok Urbančič ◽  
Christian Eggeling ◽  
Martin Booth
Keyword(s):  
Adaptive Optics ◽  
Molecular Diffusion ◽  
Three Dimensional ◽  
Super Resolution ◽  
Correction Method ◽  
Living Cells ◽  
Correlation Spectroscopy ◽  
Stimulated Emission ◽  
Optical Aberrations ◽  
Extreme Sensitivity

Fluorescence correlation spectroscopy in combination with super-resolution stimulated emission depletion microscopy (STED-FCS) is a powerful tool to investigate molecular diffusion with sub-diffraction resolution. It has been of particular use for investigations of two dimensional systems like cell membranes, but has so far seen very limited applications to studies of three-dimensional diffusion. One reason for this is the extreme sensitivity of the axial (3D) STED depletion pattern to optical aberrations. We present here an adaptive optics-based correction method that compensates for these aberrations and allows STED-FCS measurements in the cytoplasm of living cells.

scholarly journals 3D super-resolved imaging in live cells using sub-diffractive plasmonic localization of hybrid nanopillar arrays

Nanophotonics ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 2847-2859
Author(s):  
Soojung Kim ◽  
Hyerin Song ◽  
Heesang Ahn ◽  
Seung Won Jun ◽  
Seungchul Kim ◽  
...  
Keyword(s):  
Signal To Noise Ratio ◽  
Imaging Techniques ◽  
Optical Loss ◽  
Super Resolution ◽  
Living Cells ◽  
Live Cells ◽  
Complex Biological System ◽  
Observation Volume ◽  
Resolution Imaging ◽  
Nanopillar Arrays

AbstractAnalysing dynamics of a single biomolecule using high-resolution imaging techniques has been had significant attentions to understand complex biological system. Among the many approaches, vertical nanopillar arrays in contact with the inside of cells have been reported as a one of useful imaging applications since an observation volume can be confined down to few-tens nanometre theoretically. However, the nanopillars experimentally are not able to obtain super-resolution imaging because their evanescent waves generate a high optical loss and a low signal-to-noise ratio. Also, conventional nanopillars have a limitation to yield 3D information because they do not concern field localization in z-axis. Here, we developed novel hybrid nanopillar arrays (HNPs) that consist of SiO2 nanopillars terminated with gold nanodisks, allowing extreme light localization. The electromagnetic field profiles of HNPs are obtained through simulations and imaging resolution of cell membrane and biomolecules in living cells are tested using one-photon and 3D multiphoton fluorescence microscopy, respectively. Consequently, HNPs present approximately 25 times enhanced intensity compared to controls and obtained an axial and lateral resolution of 110 and 210 nm of the intensities of fluorophores conjugated with biomolecules transported in living cells. These structures can be a great platform to analyse complex intracellular environment.

scholarly journals Numerically Enhanced Stimulated Emission Depletion Microscopy with Adaptive Optics for Deep-Tissue Super-Resolved Imaging

ACS Nano ◽  
2019 ◽  
Vol 14 (1) ◽  
pp. 394-405 ◽  
Author(s):  
Piotr Zdańkowski ◽  
Maciej Trusiak ◽  
David McGloin ◽  
Jason R. Swedlow
Keyword(s):  
Adaptive Optics ◽  
Stimulated Emission ◽  
Deep Tissue ◽  
Stimulated Emission Depletion

Recent advances in luminescent materials for super-resolution imaging via stimulated emission depletion nanoscopy

2021 ◽  
Author(s):  
Yanzi Xu ◽  
Ruohan Xu ◽  
Zhi Wang ◽  
Yu Zhou ◽  
Qifei Shen ◽  
...  
Keyword(s):  
Recent Progress ◽  
Super Resolution ◽  
Stimulated Emission ◽  
Luminescent Materials ◽  
New Materials ◽  
Recent Advances ◽  
Resolution Imaging ◽  
Stimulated Emission Depletion

Recent progress on STED fluorophores for super-resolution imaging and also their characteristics are outlined here, thus providing some guidelines to select proper probes and even develop new materials for super-resolution imaging via STED nanoscopy.

Long-term STED imaging of amyloid fibers with exchangeable Thioflavin T

Nanoscale ◽  
2020 ◽  
Vol 12 (28) ◽  
pp. 15050-15053
Author(s):  
Joaquim Torra ◽  
Patricia Bondia ◽  
Sylvia Gutierrez-Erlandsson ◽  
Begoña Sot ◽  
Cristina Flors
Keyword(s):  
Super Resolution ◽  
Stimulated Emission ◽  
Thioflavin T ◽  
Amyloid Fibers ◽  
Resolution Imaging ◽  
Stimulated Emission Depletion

We report the use of the amyloid probe Thioflavin T (ThT) as a specific and exchangeable fluorophore for stimulated emission depletion (STED) super-resolution imaging of amyloid fibers.

scholarly journals PSF shaping using adaptive optics for three-dimensional single-molecule super-resolution imaging and tracking

2012 ◽  
Vol 20 (5) ◽  
pp. 4957 ◽  
Author(s):  
Ignacio Izeddin ◽  
Mohamed El Beheiry ◽  
Jordi Andilla ◽  
Daniel Ciepielewski ◽  
Xavier Darzacq ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Single Molecule ◽  
Three Dimensional ◽  
Super Resolution ◽  
Resolution Imaging

scholarly journals Adaptive optics improves multiphoton super-resolution imaging

2017 ◽  
Vol 14 (9) ◽  
pp. 869-872 ◽  
Author(s):  
Wei Zheng ◽  
Yicong Wu ◽  
Peter Winter ◽  
Robert Fischer ◽  
Damian Dalle Nogare ◽  
...  
Keyword(s):  
Adaptive Optics ◽  
Super Resolution ◽  
Resolution Imaging
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