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Label-free adaptive optics single-molecule localization microscopy for whole animals

Sanghyeon Park, Yonghyeon Jo, Minsu Kang, Jin Hee Hong, Sangyoon Ko, Suhyun Kim, Sangjun Park, Hae-Chul Park, Sang-Hee Shim, Wonshik Choi
doi: https://doi.org/10.1101/2021.11.18.469175
Sanghyeon Park
1Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science, Seoul 02841, Korea
2Department of Physics, Korea University, Seoul 02855, Korea
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Yonghyeon Jo
1Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science, Seoul 02841, Korea
2Department of Physics, Korea University, Seoul 02855, Korea
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Minsu Kang
3Department of Chemistry, Korea University, Seoul 02855, Korea
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Jin Hee Hong
1Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science, Seoul 02841, Korea
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Sangyoon Ko
3Department of Chemistry, Korea University, Seoul 02855, Korea
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Suhyun Kim
5Department of Biomedical Sciences, Korea University, Asan 425-707, Korea
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Sangjun Park
4Department of Medical Life Sciences, The Catholic University of Korea, Seoul 06591, Korea
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Hae-Chul Park
5Department of Biomedical Sciences, Korea University, Asan 425-707, Korea
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Sang-Hee Shim
3Department of Chemistry, Korea University, Seoul 02855, Korea
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Wonshik Choi
1Center for Molecular Spectroscopy and Dynamics, Institute for Basic Science, Seoul 02841, Korea
2Department of Physics, Korea University, Seoul 02855, Korea
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  • For correspondence: wonshik@korea.ac.kr
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Abstract

The specimen-induced aberration has been a major factor limiting the imaging depth of single-molecule localization microscopy (SMLM). Here, we report the application of label-free wavefront sensing adaptive optics to SMLM for deep-tissue super-resolution imaging. The proposed system measures complex tissue aberrations from intrinsic reflectance rather than fluorescence emission and physically corrects the wavefront distortion more than three-fold stronger than the previous limit. This enables us to resolve sub-diffraction morphologies of cilia and oligodendrocytes in whole intact zebrafish as well as dendritic spines in thick mouse brain tissues at the depth of up to 102 μm with localization number enhancement by up to 37 times and localization precision comparable to aberration-free samples. The proposed approach can expand the application range of SMLM to intact animals that cause the loss of localization points owing to severe tissue aberrations.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • ↵* sangheeshim{at}korea.ac.kr and wonshik{at}korea.ac.kr

  • Updated typos and mis-typed numerical values and added new data.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. All rights reserved. No reuse allowed without permission.
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Posted October 26, 2022.
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Label-free adaptive optics single-molecule localization microscopy for whole animals
Sanghyeon Park, Yonghyeon Jo, Minsu Kang, Jin Hee Hong, Sangyoon Ko, Suhyun Kim, Sangjun Park, Hae-Chul Park, Sang-Hee Shim, Wonshik Choi
bioRxiv 2021.11.18.469175; doi: https://doi.org/10.1101/2021.11.18.469175
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Label-free adaptive optics single-molecule localization microscopy for whole animals
Sanghyeon Park, Yonghyeon Jo, Minsu Kang, Jin Hee Hong, Sangyoon Ko, Suhyun Kim, Sangjun Park, Hae-Chul Park, Sang-Hee Shim, Wonshik Choi
bioRxiv 2021.11.18.469175; doi: https://doi.org/10.1101/2021.11.18.469175

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