Extended-depth of field random illumination microscopy, EDF-RIM, provides super-resolved projective imaging

Lorry Mazzella; Thomas Mangeat; Guillaume Giroussens; Benoit Rogez; Hao Li; Justine Creff; Mehdi Saadaoui; Carla Martins; Ronan Bouzignac; Simon Labouesse; Jérome Idier; Frédéric Galland; Marc Allain; Anne Sentenac; Loïc LeGoff

doi:10.1038/s41377-024-01612-0

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Extended-depth of field random illumination microscopy, EDF-RIM, provides super-resolved projective imaging

Original Articles

- Extended-depth of field random illumination microscopy, EDF-RIM, provides super-resolved projective imaging
- Light: Science & Applications Vol. 13, Issue 12, Pages: 3052-3063(2024)
- Affiliations：
  
  1.Aix Marseille Université, CNRS, Centrale Med, Institut Fresnel UMR7249, Turing Center for Living Systems, Marseille, France
  2.LITC Core Facility, Centre de Biologie Integrative (CBI), CNRS, Université de Toulouse, UT3, Toulouse, France
  3.MCD, Centre de Biologie Intégrative (CBI), CNRS, Université de Toulouse, UT3, Toulouse, France
  4.Aix Marseille University, CNRS, IBDM UMR7288, Turing Centre for Living Systems, Marseille, France
  5.LS2N, CNRS UMR 6004, F44321 Nantes Cedex 3, France
- Author bio：
  
  Anne Sentenac (anne.sentenac@fresnel.fr)
  Loïc LeGoff (loic.le-goff@univ-amu.fr)
- Funds：
- DOI：10.1038/s41377-024-01612-0
  CLC：
- Published：31 December 2024，
  
  Published Online：10 October 2024，
  
  Received：30 October 2023，
  
  Revised：15 August 2024，
  
  Accepted：27 August 2024
- Accepted：
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Mazzella, L. et al. Extended-depth of field random illumination microscopy, EDF-RIM, provides super-resolved projective imaging. Light: Science & Applications, 13, 3052-3063 (2024).
DOI：

Mazzella, L. et al. Extended-depth of field random illumination microscopy, EDF-RIM, provides super-resolved projective imaging. Light: Science & Applications, 13, 3052-3063 (2024). DOI： 10.1038/s41377-024-01612-0.

摘要

Abstract

The ultimate aim of fluorescence microscopy is to achieve high-resolution imaging of increasingly larger biological samples. Extended depth of field presents a potential solution to accelerate imaging of large samples when compression of information along the optical axis is not detrimental to the interpretation of images. We have implemented an extended depth of field (EDF) approach in a random illumination microscope (RIM). RIM uses multiple speckled illuminations and variance data processing to double the resolution. It is particularly adapted to the imaging of thick samples as it does not require the knowledge of illumination patterns. We demonstrate highly-resolved projective images of biological tissues and cells. Compared to a sequential scan of the imaged volume with conventional 2D-RIM

EDF-RIM allows an order of magnitude improvement in speed and light dose reduction

with comparable resolution. As the axial information is lost in an EDF modality

we propose a method to retrieve the sample topography for samples that are organized in cell sheets.

关键词

Keywords

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