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

Sanghyeon Park, Yonghyeon Jo, Minsu Kang, Jin Hee Hong, Sangyoon Ko, Suhyun Kim, Sangjun Park, Hae Chul Park, Sang-Hee Shim () and Wonshik Choi ()
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Sanghyeon Park: Institute for Basic Science
Yonghyeon Jo: Institute for Basic Science
Minsu Kang: Korea University
Jin Hee Hong: Institute for Basic Science
Sangyoon Ko: Korea University
Suhyun Kim: Korea University
Sangjun Park: The Catholic University of Korea
Hae Chul Park: Korea University
Sang-Hee Shim: Korea University
Wonshik Choi: Institute for Basic Science

Nature Communications, 2023, vol. 14, issue 1, 1-11

Abstract: Abstract 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 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 whole zebrafish that cause the loss of localization number owing to severe tissue aberrations.

Date: 2023
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DOI: 10.1038/s41467-023-39896-2

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