title: An integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos.
abstract: Understanding the fundamental principles governing embryogenesis is a key goal of developmental biology. Direct observation of embryogenesis via in vivo live imaging is vital to understanding embryogenesis; yet, tedious sample preparation makes it difficult to acquire large-scale imaging data that is often required to overcome experimental and biological noises for quantitative studies. Furthermore, it is often difficult, and sometimes impossible, to incorporate environmental perturbation for understanding developmental responses to external stimuli. To address this issue, we have developed a method for high-throughput imaging of live embryos, delivering precise environmental perturbations, and unbiased data extraction. This platform includes an optimized microfluidic device specifically for live embryos and also for precise perturbations in the microenvironment of the developing embryos. In addition, we developed software for simple, yet accurate, automated segmentation of fluorescent images, and automated data extraction. Using a quantitative assessment we find that embryos develop normally within the microfluidic device. Finally, we show an application of the high-throughput assay for monitoring developmental responses to external stimuli: anoxia-induced developmental arrest in Drosophila embryos. With slight modifications, the method developed in this work can be applied to many other models of development and other stimulus-response behaviors during development.
authors: School of Chemical &Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA, USA 30332.
authors: Thomas J
authors: TJ
authors: Levario
authors: Walter H Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA 30332.
authors: Charles
authors: C
authors: Zhao
authors: School of Chemical &Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA, USA 30332.
authors: Tel
authors: T
authors: Rouse
authors: Department of Chemical and Biological Engineering and Lewis-Sigler Institute for Integrative Genomics, Princeton University, Princeton, New Jersey, USA 08544.
authors: Stanislav Y
authors: SY
authors: Shvartsman
authors: School of Chemical &Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA, USA 30332.|Walter H Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA, USA 30332.
authors: Hang
authors: H
authors: Lu
mesh_terms: D000818:Animals; D004331:Drosophila melanogaster / Q000196:embryology* / Q000648:ultrastructure; D004625:Embryo, Nonmammalian; D047108:Embryonic Development / Q000502:physiology*; D057166:High-Throughput Screening Assays; D000860:Hypoxia / Q000473:pathology; D007091:Image Processing, Computer-Assisted; D056656:Lab-On-A-Chip Devices; D061848:Optical Imaging / Q000295:instrumentation* / Q000379:methods*; D012984:Software