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. 2018 Feb 23:(132):56580.
doi: 10.3791/56580.

Lens-free Video Microscopy for the Dynamic and Quantitative Analysis of Adherent Cell Culture

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Lens-free Video Microscopy for the Dynamic and Quantitative Analysis of Adherent Cell Culture

Cedric Allier et al. J Vis Exp. .

Abstract

Here, we demonstrate that lens-free video microscopy enables us to simultaneously capture the kinetics of thousands of cells directly inside the incubator and that it is possible to monitor and quantify single cells along several cell cycles. We describe the full protocol used to monitor and quantify a HeLa cell culture for 2.7 days. First, cell culture acquisition is performed with a lens-free video microscope, and then the data is analyzed following a four-step process: multi-wavelength holographic reconstruction, cell-tracking, cell segmentation and cell division detection algorithms. As a result, we show that it is possible to gather a dataset featuring more than 10,000 cell cycle tracks and more than 2 x 106 cell morphological measurements.

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