In-depth single molecule localization microscopy using adaptive optics and single objective light-sheet microscopy

Marine Cabillic, Hisham Forriere, Laetitia Bettarel, Corey Butler, Abdelghani Neuhaus, Ihssane Idrissi, Miguel Edouardo Sambrano-Lopez, Julian Rossbroich, Lucas-Raphael Müller, Jonas Ries, Gianluca Grenci, Virgile Viasnoff, Florian Levet, Jean-Baptiste Sibarita () and Rémi Galland ()
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Marine Cabillic: Univ. Bordeaux, CNRS, IINS, UMR 5297
Hisham Forriere: Univ. Bordeaux, CNRS, IINS, UMR 5297
Laetitia Bettarel: Univ. Bordeaux, CNRS, IINS, UMR 5297
Corey Butler: Univ. Bordeaux, CNRS, IINS, UMR 5297
Abdelghani Neuhaus: Univ. Bordeaux, CNRS, IINS, UMR 5297
Ihssane Idrissi: Univ. Bordeaux, CNRS, IINS, UMR 5297
Miguel Edouardo Sambrano-Lopez: Univ. Bordeaux, CNRS, IINS, UMR 5297
Julian Rossbroich: Univ. Bordeaux, CNRS, IINS, UMR 5297
Lucas-Raphael Müller: European Molecular Biology Laboratory (EMBL)
Jonas Ries: European Molecular Biology Laboratory (EMBL)
Gianluca Grenci: National University of Singapore
Virgile Viasnoff: National University of Singapore
Florian Levet: Univ. Bordeaux, CNRS, IINS, UMR 5297
Jean-Baptiste Sibarita: Univ. Bordeaux, CNRS, IINS, UMR 5297
Rémi Galland: Univ. Bordeaux, CNRS, IINS, UMR 5297

Nature Communications, 2025, vol. 16, issue 1, 1-15

Abstract: Abstract Single molecule localization microscopy (SMLM) allows deciphering the nanoscale organization and dynamics of biomolecules in their native environment with unprecedented resolution. While SMLM was quickly adopted by the scientific community for its performance and simple instrumentation, it still remains limited in its in-depth capability, precluding many biological processes to be investigated. We here present a solution to perform in-depth volumetric SMLM, called soSMARt. It relies on innovative microfabricated devices allowing both single-objective light-sheet microscopy, aberrations correction via adaptive optics, and real-time feedback-loop registration with nanometric precision. We illustrated the performances of soSMARt to assess the 3D nanoscale organization of several protein of interest in isolated cells, and explore optimizations and proof-of-concepts for the investigation of more complex tissues such as 3D cell cultures. We believe our method addresses key limitations of single molecule microscopy, paving the way for novel biological applications.

Date: 2025
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DOI: 10.1038/s41467-025-62198-8

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