Lensfree diffractive tomography for the imaging of 3D cell cultures
Résumé
New microscopes are needed to help realize the full potential of 3D organoid culture studies by
gathering large quantitative and systematic data over extended period of time while preserving the integrity
of the living sample. On another hand, lensfree video microscopy is addressing these needs in the context of
2D cell culture, providing label-free and non-phototoxic acquisition of large datasets. As scientists
routinely adopt 3D culture techniques, the new challenging task is to extend lensfree microscopy techniques
to 3D structures.
In order to image such large volume while preserving the ability to catch every single cell, we propose
new imaging platforms based on lensfree microscopy. We have built lensfree diffractive tomography setups
performing multi-angle acquisitions of 3D organoid culture embedded in Matrigel® and developed a
dedicated 3D holographic reconstruction algorithm based on the Fourier diffraction theorem.
With a first prototype we have been able to reconstruct a 3D volume as large as 21.5 mm^3 of a 3D
organoid culture of prostatic RWPE1 cells in Matrigel® showing the ability of these cells to assemble in 3D
intricate cellular network at the mesoscopic scale. Comparisons with 2D images showed that it is
possible to detect single cells isolated from the main cellular structure.
From results of the first prototype and numerical simulations, a second prototype was
designed to improve the reconstructions quality, especially on the vertical axis. First results on dust and
bubbles lying at the bottom of a Petri® dish are promising.
The adaptation of lensfree microscopy techniques to 3D organoid cultures imaging and the
presentation of the associated algorithms are the scopes of the present communication.
Domaines
Optique / photonique
Origine : Fichiers produits par l'(les) auteur(s)