Ms. Anaïs Saintoyant Profile

Anaïs Saintoyant

at PHASICS SA

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Author

Publications (3)

Proceedings Article | 11 March 2020 Presentation

Non-invasive intracellular dynamics study using quadriwave lateral shearing interferometry (Conference Presentation)

Roman Zinchuk, Julien Savatier, Anaïs Saintoyant, Sherazade Aknoun, Antoine Federici, Serge Monneret, Benoit Wattellier

Proceedings Volume 11249, 112490W (2020) https://doi.org/10.1117/12.2544711

KEYWORDS: Interferometry, Algorithm development, Phase imaging, Cytoskeletons, Detection and tracking algorithms, Molecules, Microscopy, Image resolution, Wavefront sensors, Detector development

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Proceedings Article | 4 March 2019 Presentation

Semi-automated imaging station dedicated to quantitative phase imaging for the study of large numbers of cells over long period of time (Conference Presentation)

Anaïs Saintoyant, Sherazade Aknoun, Antoine Federici, Fabrice Valentino, Benoit Wattellier

Proceedings Volume 10887, 1088716 (2019) https://doi.org/10.1117/12.2509739

KEYWORDS: Phase imaging, Image processing, Algorithm development, Interferometry, Data processing, Data acquisition, Microscopes, Image acquisition, Natural surfaces, Tissues

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High content screening consists in acquiring a large number of samples to obtain statistically significant information on cell populations and their changes over time. It is also used to compare different growth conditions. Quantitative phase imaging enables to image semi-transparent samples without any label. This technology has the advantage of not modifying samples so as not to disturb them and to study them over a long period of time (> 3 days) by providing relevant quantitative phase information. We develop a solution to follow a very large number of cells (> 1000) over a long period of time (> 72 h). We set up a semi-automated imaging station using QuadriWave Lateral Shearing Interferometry. This station solves at least four challenges: - Scan an entire multi-well plate within minutes - Keep focus over time and distance at high magnification - Correct for meniscus effect at the well edges - Process data as fast as it is acquired To image wide fields, sample is scanned thanks to the microscope stage. Work on synchronization and analysis’ speed with GPU made it possible to perform such a scan at a speed making possible the cell follow-up between two acquisitions. As scanned surface is large and time lapse can be performed over long periods of time, a numerical refocusing algorithm processes phase images even after acquisition. Performances and limitations of this approach will be presented. Different algorithms developed to stitch phase images together enable to get quantitative phase information over the whole field of view. With this system, we perform scans of entire surface of tissue samples with high resolution. We are also able to image entire wells of multiwell plates with phase modality. It is then possible to study the evolution of quantitative phase and morphological features for long time periods at the individual cell level for a large number of cells evolving in different wells according to different conditions. We will present cell growth comparison for different experimental culture conditions.

Proceedings Article | 4 March 2019 Presentation

Intracellular dynamics with quantitative phase imaging (Conference Presentation)

Sherazade Aknoun, Anaïs Saintoyant, Antoine Federici, Benoit Wattellier

Proceedings Volume 10887, 108871K (2019) https://doi.org/10.1117/12.2510536

KEYWORDS: Phase imaging, Molecules, Cytoskeletons, Cell biology, Biology, Proteins, Luminescence, Microscopy, Image resolution, Wavefront sensors

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It is now known that interaction between cells and their environment or between intracellular compartments is based on complex vesicular transport processes [1]. This transport consists in material internalization from the external environment and compartment exchanges inside the cell itself. Understanding the mechanisms and regulation of this intracellular trafficking is an intense object of study in the field of cellular biology. The goal is to understand how vesicles transporting proteins and lipids are targeted to specific cellular compartments and fused with the membrane. Progress about intracellular trafficking is currently essentially made by constant innovation in fluorescence based techniques. They now reach single molecule resolution in living cells. It is possible to follow molecules all along their travel inside the cell. In this case, the main limitation is fluorescent probes could bias the vesicle behaviors and alter their transport inside the cell. Quantitative phase imaging techniques are conventionally used in microscopy, enhancing the contrast for imaging semi-transparent samples with a non-invasive (i.e. label free) and fast approach. For instance, phase correlation imaging introduced by [2] showed that global statistics on the whole cell or a population can give insight of motions and dynamics. We propose to study in details the behavior of vesicles at the intracellular scale and their interaction with the cytoskeleton. The resolution and the high sensitivity brought by a High Definition wave front sensor allows following all individual vesicles through time follow up, while imaging the whole cell. We can study their interaction with the surrounding environment (Brownian or guided motion along the cytoskeleton). We will show how we can extract information from quantitative phase images about the intracellular transport and the influence of the intracellular order/disorder. [1] Tokarev AA, Alfonso A, Segev N. Overview of Intracellular Compartments and Trafficking Pathways. In: Madame Curie Bioscience Database [Internet]. Austin (TX): Landes Bioscience; 2000-2013. [2] Phase correlation imaging of unlabeled cell dynamics, Lihong Ma and al., Scientific Reports volume 6, Article number: 32702 (2016)

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