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Self-interference 3D super-resolution microscopy for deep tissue investigations.

Pierre Bon1,2, Jeanne Linarès-Loyez3,4, Maxime Feyeux3,4

  • 1Université de Bordeaux, Laboratoire Photonique Numérique et Nanosciences, UMR 5298, F-33400, Talence, France. pierre.bon@u-bordeaux.fr.

Nature Methods
|May 2, 2018
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Summary
This summary is machine-generated.

We developed SELFI, a novel 3D single-molecule localization microscopy technique. This method enables nanoscale imaging within intact biological tissues, overcoming previous depth limitations for cellular biology research.

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Area of Science:

  • Biophysics
  • Cellular Biology
  • Microscopy

Background:

  • Fluorescence localization microscopy offers near-molecular resolution for ultra-structure visualization in cellular biology.
  • Achieving 3D resolution within biological tissues beyond superficial layers remains a significant challenge.

Purpose of the Study:

  • To introduce a framework for 3D single-molecule localization within multicellular specimens and tissues.
  • To enable nanoscale investigations of cellular processes in intact tissues.

Main Methods:

  • Developed the SELFI (Self-interference for Localization of Fluorescence Intensity) framework.
  • Utilized self-interference within the microscope's point spread function (PSF) to encode 3D position information.
  • Combined SELFI with conventional localization microscopy techniques.

Main Results:

  • Successfully visualized 3D F-actin filament networks.
  • Revealed the spatial distribution of the transcription factor OCT4 in human induced pluripotent stem cells.
  • Achieved imaging at depths up to 50 µm within uncleared tissue spheroids.

Conclusions:

  • SELFI provides a robust method for 3D single-molecule localization in intact biological tissues.
  • The framework overcomes depth limitations, enabling nanoscale imaging deeper within specimens.
  • SELFI facilitates the study of native cellular processes in their native tissue environment.