1. Dynamics of cell and tissue growth acquired by means of extended field of view lensfree microscopy
- Author
-
Jean-Guillaume Coutard, Thomas Bordy, Cédric Allier, Fabrice Navarro, Mathilde Menneteau, Fabien Momey, J.-M. Dinten, Commissariat à l'énergie atomique et aux énergies alternatives - Laboratoire d'Electronique et de Technologie de l'Information (CEA-LETI), Direction de Recherche Technologique (CEA) (DRT (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Laboratoire Hubert Curien [Saint Etienne] (LHC), Institut d'Optique Graduate School (IOGS)-Université Jean Monnet [Saint-Étienne] (UJM)-Centre National de la Recherche Scientifique (CNRS), Ferme expérimentale de Thorigné d'Anjou, Centre de Recherche en Economie et Statistique [Bruz] (CREST), Ecole Nationale de la Statistique et de l'Analyse de l'Information [Bruz] (ENSAI), Laboratoire Hubert Curien (LHC), and Institut d'Optique Graduate School (IOGS)-Université Jean Monnet - Saint-Étienne (UJM)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
0301 basic medicine ,CMOS sensor ,Computer science ,business.industry ,Image processing ,Video microscopy ,Field of view ,Velocimetry ,Atomic and Molecular Physics, and Optics ,Article ,03 medical and health sciences ,030104 developmental biology ,Optics ,[INFO.INFO-IM]Computer Science [cs]/Medical Imaging ,Computer vision ,Artificial intelligence ,Image sensor ,Raster scan ,business ,Digital holography ,ComputingMilieux_MISCELLANEOUS ,Biotechnology - Abstract
In this paper, we discuss a new methodology based on lensfree imaging to perform wound healing assay with unprecedented statistics. Our video lensfree microscopy setup is a simple device featuring only a CMOS sensor and a semi coherent illumination system. Yet it is a powerful mean for the real-time monitoring of cultivated cells. It presents several key advantages, e.g. integration into standard incubator, compatibility with standard cell culture protocol, simplicity and ease of use. It can perform the follow-up in a large field of view (25 mm(2)) of several crucial parameters during the culture of cells i.e. their motility, their proliferation rate or their death. Consequently the setup can gather large statistics both in space and time. Here we uses this facility in the context of wound healing assay to perform label-free measurements of the velocities of the fronts of proliferation of the cell layer as a function of time by means of particle image velocimetry (PIV) processing. However, for such tissue growth experiments, the field of view of 25 mm(2) remains not sufficient and results can be biased depending on the position of the device with respect to the recipient of the cell culture. Hence, to conduct exhaustive wound healing assays, we propose to enlarge the field of view up to 10 cm(2) through a raster scan, by moving the source/sensor with respect to the Petri dish. We have performed acquisitions of wound healing assay (keratinocytes HaCaT) both in real-time (25 mm(2)) and in final point (10 cm(2)) to assess the combination of velocimetry measurements and final point wide field imaging. In the future, we aim at combining directly our extended field of view acquisitions (10 cm(2)) with real time ability inside the incubator.
- Published
- 2015
- Full Text
- View/download PDF