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

Piotr Zdańkowski; Maciej Trusiak; David McGloin; Jason R. Swedlow

doi:10.1021/acsnano.9b05891

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

Piotr Zdańkowski, Maciej Trusiak, David McGloin, Jason R. Swedlow^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

19 Citations (Scopus)

Abstract

In 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 utilizing a high-power depletion laser (increasing the risk of photodamage), while the depletion beam is very sensitive to sample-induced aberrations. Here, we demonstrate a custom-built STED microscope with automated aberration correction that is capable of 3D super-resolution imaging through thick, highly aberrating tissue. We introduce and investigate a state of the art image denoising method by block-matching and collaborative 3D filtering (BM3D) to numerically enhance fine object details otherwise mixed with noise and further enhance the image quality. Numerical denoising provides an increase in the final effective resolution of the STED imaging of 31% using the well established Fourier ring correlation metric. Results achieved through the combination of aberration correction and tailored image processing are experimentally validated through super-resolved 3D imaging of axons in differentiated induced pluripotent stem cells growing under an 80 μm thick layer of tissue with lateral and axial resolution of 204 and 310 nm, respectively.

Original language	English
Pages (from-to)	394-405
Number of pages	12
Journal	ACS Nano
Volume	14
Issue number	1
Early online date	16 Dec 2019
DOIs	https://doi.org/10.1021/acsnano.9b05891
Publication status	Published - 28 Jan 2020

Bibliographical note

Funding Information:
This work was funded by the People Programme (Marie Curie Actions) of the European Union’s Seventh Framework Programme (FP7/2007-2013) under REA grant agreement no. 608133. The microscope development was supported by the Wolfson Foundation and the Scottish Universities Physics Alliance (SUPA). P.Z. and M.T. would like to acknowledge the support of the National Science Center, Poland, under the project OPUS 13 (UMO2017/25/B/ST7/02049), National Agency for Academic Exchange, and statutory funds of the Faculty of Mechatronics, Warsaw University of Technology. We thank Iain Porter, Michael Porter, and Lindsay Davidson for the preparation of the differentiated hiPSC derived neurons.

Publisher Copyright:
Copyright © 2019 American Chemical Society.

Keywords

aberration correction
adaptive optics
deep imaging
fluorescence microscopy
STED microscopy
super-resolution microscopy

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title = "Numerically Enhanced Stimulated Emission Depletion Microscopy with Adaptive Optics for Deep-Tissue Super-Resolved Imaging",

abstract = "In 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 utilizing a high-power depletion laser (increasing the risk of photodamage), while the depletion beam is very sensitive to sample-induced aberrations. Here, we demonstrate a custom-built STED microscope with automated aberration correction that is capable of 3D super-resolution imaging through thick, highly aberrating tissue. We introduce and investigate a state of the art image denoising method by block-matching and collaborative 3D filtering (BM3D) to numerically enhance fine object details otherwise mixed with noise and further enhance the image quality. Numerical denoising provides an increase in the final effective resolution of the STED imaging of 31% using the well established Fourier ring correlation metric. Results achieved through the combination of aberration correction and tailored image processing are experimentally validated through super-resolved 3D imaging of axons in differentiated induced pluripotent stem cells growing under an 80 μm thick layer of tissue with lateral and axial resolution of 204 and 310 nm, respectively.",

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author = "Piotr Zda{\'n}kowski and Maciej Trusiak and David McGloin and Swedlow, {Jason R.}",

note = "Funding Information: This work was funded by the People Programme (Marie Curie Actions) of the European Union{\textquoteright}s Seventh Framework Programme (FP7/2007-2013) under REA grant agreement no. 608133. The microscope development was supported by the Wolfson Foundation and the Scottish Universities Physics Alliance (SUPA). P.Z. and M.T. would like to acknowledge the support of the National Science Center, Poland, under the project OPUS 13 (UMO2017/25/B/ST7/02049), National Agency for Academic Exchange, and statutory funds of the Faculty of Mechatronics, Warsaw University of Technology. We thank Iain Porter, Michael Porter, and Lindsay Davidson for the preparation of the differentiated hiPSC derived neurons. Publisher Copyright: Copyright {\textcopyright} 2019 American Chemical Society.",

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AB - In 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 utilizing a high-power depletion laser (increasing the risk of photodamage), while the depletion beam is very sensitive to sample-induced aberrations. Here, we demonstrate a custom-built STED microscope with automated aberration correction that is capable of 3D super-resolution imaging through thick, highly aberrating tissue. We introduce and investigate a state of the art image denoising method by block-matching and collaborative 3D filtering (BM3D) to numerically enhance fine object details otherwise mixed with noise and further enhance the image quality. Numerical denoising provides an increase in the final effective resolution of the STED imaging of 31% using the well established Fourier ring correlation metric. Results achieved through the combination of aberration correction and tailored image processing are experimentally validated through super-resolved 3D imaging of axons in differentiated induced pluripotent stem cells growing under an 80 μm thick layer of tissue with lateral and axial resolution of 204 and 310 nm, respectively.

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

Abstract

Bibliographical note

Keywords

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