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Proceedings Paper

In vivo microdissection and live embryo imaging by two-photon microscopy to study Drosophila melanogaster early development
Author(s): Willy Supatto; Eric Brouzes; Emmanuel Farge; Emmanuel Beaurepaire
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Paper Abstract

Animal embryo development exhibits a complex choreography of cell movements highly regulated both in time and space. This sequence of morphogenetic movements is initiated at gastrulation and is tightly controlled by a cascade of developmental gene expression. We have recently reported that developmental gene expression can in turn be mechanically regulated by morphogenetic movements during Drosophila melanogaster early development. In order to study this phenomenon of mechanically induced gene expression, it is necessary to develop new techniques of in vivo investigation. We show that the combination of femtosecond pulse intratissue surgery and two-photon-excitation fluorescence (2PEF) microscopy is a powerful tool for (i) disrupting natural morphogenetic movements and (ii) imaging native and disrupted morphogenetic movements during Drosophila development. (i) First, non-linear-absorption-mediated photo-disruption makes it possible to perform controlled intra-vital micro-dissections resulting in the modulation of morphogenetic movements and subsequent mechano-sensitive gene expression. (ii) Second, in vivo 2PEF microscopy of transgenic GFP systems appears to be an excellent technique for long-term in vivo imaging of the complex morphogenetic movements involved in normal or perturbed Drosophila gastrulation. Together, these two techniques provide a powerful novel approach to study embryo development.

Paper Details

Date Published: 8 September 2004
PDF: 8 pages
Proc. SPIE 5463, Femtosecond Laser Applications in Biology, (8 September 2004); doi: 10.1117/12.545453
Show Author Affiliations
Willy Supatto, Institut Curie, CNRS (France)
Univ. Paris VII (France)
Eric Brouzes, Institut Curie, CNRS (France)
Emmanuel Farge, Institut Curie, CNRS (France)
Emmanuel Beaurepaire, Lab. for Optics and Biosciences, CNRS, INSERM, and Ecole Polytechnique (France)

Published in SPIE Proceedings Vol. 5463:
Femtosecond Laser Applications in Biology
Sigrid Avrillier; Jean-Michel Tualle, Editor(s)

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