An integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos.

TitleAn integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos.
Publication TypeJournal Article
Year of Publication2016
AuthorsLevario, TJ, Zhao, C, Rouse, T, Shvartsman, SY, Lu, H
JournalSci Rep
Volume6
Pagination21366
Date Published2016 Feb 11
ISSN2045-2322
KeywordsAnimals, Drosophila melanogaster, Embryo, Nonmammalian, Embryonic Development, High-Throughput Screening Assays, Hypoxia, Image Processing, Computer-Assisted, Lab-On-A-Chip Devices, Optical Imaging, Software
Abstract

<p>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. </p>

DOI10.1038/srep21366
Alternate JournalSci Rep
PubMed ID26864815
PubMed Central IDPMC4750044
Grant ListGM078079 / GM / NIGMS NIH HHS / United States
R01 GM078079 / GM / NIGMS NIH HHS / United States
R01GM086537 / GM / NIGMS NIH HHS / United States
GM088333 / GM / NIGMS NIH HHS / United States
R01 GM088333 / GM / NIGMS NIH HHS / United States
1F31AA023160-01 / AA / NIAAA NIH HHS / United States
R01 GM086537 / GM / NIGMS NIH HHS / United States
F31 AA023160 / AA / NIAAA NIH HHS / United States