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Journal of Biomedical Optics • Open Access

Motility flow and growth-cone navigation analysis during in vitro neuronal development by long-term bright-field imaging
Author(s): Maya Shalev Aviv; Mattia Pesce; Sharada Tilve; Evelina Chieregatti; Zeev Zalevsky; Francesco Difato

Paper Abstract

A long-term live-imaging workstation to follow the development of cultured neurons during the first few days in vitro (DIV) is developed. In order to monitor neuronal polarization and axonal growth by live imaging, we built a micro-incubator system that provides stable temperature, pH, and osmolarity in the culture dish under the microscope, while preserving environment sterility. We are able to image living neurons at 2 DIVs for 48 h with a temporal resolution of one frame for every 2 min. The main features of this system are its ability to adapt to every cell-culture support, to integrate in any optical microscope, because of the relatively small dimensions (9.5×6.5×2.5  cm ) and low weight of the system (<200  g ), and to monitor the physiological parameters in situ. Moreover, we developed an image-analysis algorithm to quantify the cell motility, in order to characterize its complex temporal-spatial pattern. The algorithm applies morphological image processing operations on the temporal variations occurring in the inspected region of interest. Here, it is used to automatically detect cellular motility in three distinct morphological regions of the neurons: around the soma, along the neurites, and in the growth cone.

Paper Details

Date Published: 20 September 2013
PDF: 9 pages
J. Biomed. Opt. 18(11) 111415 doi: 10.1117/1.JBO.18.11.111415
Published in: Journal of Biomedical Optics Volume 18, Issue 11
Show Author Affiliations
Maya Shalev Aviv, Bar-Ilan Univ. (Israel)
Mattia Pesce
Sharada Tilve
Evelina Chieregatti, Istituto Italiano di Tecnologia (Italy)
Zeev Zalevsky, Bar-Ilan Univ. (Israel)
Francesco Difato, Istituto Italiano di Tecnologia (Italy)

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