TY - JOUR
T1 - An integrated platform for large-scale data collection and precise perturbation of live Drosophila embryos
AU - Levario, Thomas J.
AU - Zhao, Charles
AU - Rouse, Tel
AU - Shvartsman, Stanislav Y.
AU - Lu, Hang
N1 - Funding Information:
This work was supported by the US National Science Foundation (EFRI 1136913 to S.Y.S. and H.L.), and the US National Institutes of Health (GM078079 and R01GM086537 to S.Y.S., 1F31AA023160-01 to T.J.L. and GM088333 to H.L.). The authors thank Dr. Bomyi Lim and graduate student Yonghyun Song for technical assistance, and useful discussions. T.J.L. thanks Andrew Shaw at the Parker H. Petit Institute for Bioengineering and Bioscience microscopy core for technical assistance with confocal microscopy.
PY - 2016/2/11
Y1 - 2016/2/11
N2 - 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.
AB - 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.
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U2 - 10.1038/srep21366
DO - 10.1038/srep21366
M3 - Article
C2 - 26864815
AN - SCOPUS:84958568767
SN - 2045-2322
VL - 6
JO - Scientific reports
JF - Scientific reports
M1 - 21366
ER -