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

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

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 i...

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Bibliographic Details
Published in:Light, science & applications Vol. 13; no. 1; pp. 285 - 12
Main Authors: Mazzella, Lorry, Mangeat, Thomas, Giroussens, Guillaume, Rogez, Benoit, Li, Hao, Creff, Justine, Saadaoui, Mehdi, Martins, Carla, Bouzignac, Ronan, Labouesse, Simon, Idier, Jérome, Galland, Frédéric, Allain, Marc, Sentenac, Anne, LeGoff, Loïc
Format: Journal Article
Language:English
Published: London Nature Publishing Group UK 10-10-2024
Springer Nature B.V
Nature Publishing Group
Subjects:
639/624/1107/328/2236
639/624/1107/328/2238
Fluorescence microscopy
Illumination
Lasers
Microscopy
Microwaves
Optical and Electronic Materials
Optical Devices
Optics
Photonics
Physics
Physics and Astronomy
RF and Optical Engineering
Super-resolution microscopy
Wide-field fluorescence microscopy
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Summary: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.
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ISSN:2047-7538
2095-5545
2047-7538
DOI:10.1038/s41377-024-01612-0