Ms. Sabine Bardin Profile

Sabine Bardin

at Institut Curie

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Proceedings Article | 30 March 2020 Presentation

Video lens-free microscopy of human cells: from standard 2D to 3D organoids culture (Conference Presentation)

Cédric Allier, Sophie Morel, Anthony Berdeu, Lionel Hervé, Thomas Bordy, Michel Bornens, Sabine Bardin, Xavier Gidrol, Nathalie Picollet-D'hahan, Sophie Morales

Proceedings Volume 11351, 113510N (2020) https://doi.org/10.1117/12.2555777

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Research is continuously developing imaging methods to better understand the structure and function of biological systems. In this paper, we describe our work to develop lens-free microscopy as a novel mean to observe and quantify cells in 2D and 3D cell culture conditions. At first, we developed a lens-free video microscope based on multiple wavelength acquisitions to perform time-lapse 2D imaging of dense cell culture inside the incubator. We demonstrated that novel phase retrieval techniques enable imaging thin cell samples with high concentration (~15000 cells over a large field of view of 29.4 mm2). The experimental data can next be further analyzed with existing cell profiling and tracking algorithms. As an example, we showed that a 7 days acquisition of a culture of HeLa cells leads to a dataset featuring 2.106 cell point measurements and 104 cell cycle tracks. Recently, we extended our work to the video-microscopy of 3D organoids culture. We showed the capability of lens-free microscopy to perform 3D+time acquisitions of 3D organoids culture. To our knowledge, our technique is the only one able to reconstruct very large volumes of 3D cell culture (~5 mm3) by phase contrast imaging. This new mean of microscopy allowed us to observe a broad range of phenomena present in 3D environments, e.g. self-organizations, displacement of large clusters, merging and interconnection over long distances (>1 mm). In addition, this 3D microscope can capture the interactions of single cells and organoids with their 3D environment, e.g. traction forces generated by large cell aggregates over long distances, up to 1.5 mm. Overall, lens-free microscopy techniques favor ease of use and label-free experimentations as well as time-lapse acquisitions of large datasets. Importantly, we consider that these lens-free microscopy technique can thus expand the repertoire of phenomena that can be studied within 2D and 3D organoids cultures.

Proceedings Article | 23 February 2018 Paper

Dynamic quantitative analysis of adherent cell cultures by means of lens-free video microscopy

C. Allier, R. Vincent, F. Navarro, M. Menneteau, L. Ghenim, X. Gidrol, T. Bordy, L. Hervé, O. Cioni, S. Bardin, M. Bornens, Y. Usson, S. Morales

Proceedings Volume 10503, 105031R (2018) https://doi.org/10.1117/12.2289525

KEYWORDS: Video microscopy, Digital holography, Microscopy, Reconstruction algorithms, Quantitative analysis

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We present our implementation of lens-free video microscopy setup for the monitoring of adherent cell cultures. We use a multi-wavelength LED illumination together with a dedicated holographic reconstruction algorithm that allows for an efficient removal of twin images from the reconstructed phase image for densities up to those of confluent cell cultures (>500 cells/mm²). We thereby demonstrate that lens-free video microscopy, with a large field of view (~30 mm²) can enable us to capture the images of thousands of cells simultaneously and directly inside the incubator. It is then possible to trace and quantify single cells along several cell cycles. We thus prove that lens-free microscopy is a quantitative phase imaging technique enabling estimation of several metrics at the single cell level as a function of time, for example the area, dry mass, maximum thickness, major axis length and aspect ratio of each cell. Combined with cell tracking, it is then possible to extract important parameters such as the initial cell dry mass (just after cell division), the final cell dry mass (just before cell division), the average cell growth rate, and the cell cycle duration. As an example, we discuss the monitoring of a HeLa cell cultures which provided us with a data-set featuring more than 10 000 cell cycle tracks and more than 2x10⁶ cell morphological measurements in a single time-lapse.

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