Super-resolved live imaging of thick biological samples with 3D Random Illumination Microscopy (3D-RIM)

Thomas Mangeat; Lorry Mazzella; Benoît Rogez; Guillaume Giroussens; Xiangyi Li; Mathilde Bernard; Pablo Vargas; Marc Allain; Simon Labouesse; Jérôme Idier; Loïc LeGoff; Anne Sentenac

doi:10.1101/2025.08.06.668879

BioengineeringbioRxivAug 6, 2025

Preprint

Super-resolved live imaging of thick biological samples with 3D Random Illumination Microscopy (3D-RIM)

Thomas Mangeat iD, Lorry Mazzella, Benoît Rogez iD, Guillaume Giroussens iD, Xiangyi Li, Mathilde Bernard iD, Pablo Vargas iD, Marc Allain iD, Simon Labouesse iD, Jérôme Idier iD, Loïc LeGoff iD, Anne Sentenac iD

Abstract

Super-resolved volume imaging of thick, live specimens is greatly hampered by sample induced aberrations and out-of-focus fluorescence. In this work, using a combination of speckled illuminations, remote focusing, three-dimensional photon reassignement and variance processing, we obtained volume images with super-resolution (110 nm transverse, 270 nm axial) on thin samples and maintained a high contrast and high resolution level throughout tens of microns of highly aberrant tissues and up to hundreds of microns in collagen scaffolds.

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