Microfluidic Trap Array for Massively Parallel Imaging of Drosophila Embryos

Overview

Journal Nat Protoc

Specialties Biology
Pathology
Science

Date 2013 Mar 16

PMID 23493069

Citations 28

Authors

Thomas J Levario

Mei Zhan

Bomyi Lim

Stanislav Y Shvartsman

Hang Lu

Affiliations

Soon will be listed here.

Abstract

Here we describe a protocol for the fabrication and use of a microfluidic device to rapidly orient >700 Drosophila embryos in parallel for end-on imaging. The protocol describes master microfabrication (∼1 d), polydimethylsiloxane molding (few hours), system setup and device operation (few minutes) and imaging (depending on application). Our microfluidics-based approach described here is one of the first to facilitate rapid orientation for end-on imaging, and it is a major breakthrough for quantitative studies on Drosophila embryogenesis. The operating principle of the embryo trap is based on passive hydrodynamics, and it does not require direct manipulation of embryos by the user; biologists following the protocol should be able to repeat these procedures. The compact design and fabrication materials used allow the device to be used with traditional microscopy setups and do not require specialized fixtures. Furthermore, with slight modification, this array can be applied to the handling of other model organisms and oblong objects.

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References

Kim Y, Andreu M, Lim B, Chung K, Terayama M, Jimenez G . Gene regulation by MAPK substrate competition. Dev Cell. 2011; 20(6):880-7. PMC: 3580161. DOI: 10.1016/j.devcel.2011.05.009. View

Reeves G, Trisnadi N, Truong T, Nahmad M, Katz S, Stathopoulos A . Dorsal-ventral gene expression in the Drosophila embryo reflects the dynamics and precision of the dorsal nuclear gradient. Dev Cell. 2012; 22(3):544-57. PMC: 3469262. DOI: 10.1016/j.devcel.2011.12.007. View

Samara C, Rohde C, Gilleland C, Norton S, Haggarty S, Yanik M . Large-scale in vivo femtosecond laser neurosurgery screen reveals small-molecule enhancer of regeneration. Proc Natl Acad Sci U S A. 2010; 107(43):18342-7. PMC: 2972960. DOI: 10.1073/pnas.1005372107. View

Kanodia J, Rikhy R, Kim Y, Lund V, Delotto R, Lippincott-Schwartz J . Dynamics of the Dorsal morphogen gradient. Proc Natl Acad Sci U S A. 2009; 106(51):21707-12. PMC: 2799810. DOI: 10.1073/pnas.0912395106. View

Witzberger M, Fitzpatrick J, Crowley J, Minden J . End-on imaging: a new perspective on dorsoventral development in Drosophila embryos. Dev Dyn. 2008; 237(11):3252-9. PMC: 2881697. DOI: 10.1002/dvdy.21752. View

Crane M, Chung K, Lu H . Computer-enhanced high-throughput genetic screens of C. elegans in a microfluidic system. Lab Chip. 2009; 9(1):38-40. DOI: 10.1039/b813730g. View

Liberman L, Reeves G, Stathopoulos A . Quantitative imaging of the Dorsal nuclear gradient reveals limitations to threshold-dependent patterning in Drosophila. Proc Natl Acad Sci U S A. 2009; 106(52):22317-22. PMC: 2799780. DOI: 10.1073/pnas.0906227106. View

Keller P, Schmidt A, Wittbrodt J, Stelzer E . Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy. Science. 2008; 322(5904):1065-9. DOI: 10.1126/science.1162493. View

Steward R . Relocalization of the dorsal protein from the cytoplasm to the nucleus correlates with its function. Cell. 1989; 59(6):1179-88. DOI: 10.1016/0092-8674(89)90773-3. View

10.

Markow T, Beall S, Matzkin L . Egg size, embryonic development time and ovoviviparity in Drosophila species. J Evol Biol. 2008; 22(2):430-4. DOI: 10.1111/j.1420-9101.2008.01649.x. View

11.

Akagi J, Khoshmanesh K, Evans B, Hall C, Crosier K, Cooper J . Miniaturized embryo array for automated trapping, immobilization and microperfusion of zebrafish embryos. PLoS One. 2012; 7(5):e36630. PMC: 3351474. DOI: 10.1371/journal.pone.0036630. View

12.

Tomer R, Khairy K, Amat F, Keller P . Quantitative high-speed imaging of entire developing embryos with simultaneous multiview light-sheet microscopy. Nat Methods. 2012; 9(7):755-63. DOI: 10.1038/nmeth.2062. View

13.

Chung K, Rivet C, Kemp M, Lu H . Imaging single-cell signaling dynamics with a deterministic high-density single-cell trap array. Anal Chem. 2011; 83(18):7044-52. PMC: 3190639. DOI: 10.1021/ac2011153. View

14.

Bier E . Drosophila, the golden bug, emerges as a tool for human genetics. Nat Rev Genet. 2005; 6(1):9-23. DOI: 10.1038/nrg1503. View

15.

Chung K, Crane M, Lu H . Automated on-chip rapid microscopy, phenotyping and sorting of C. elegans. Nat Methods. 2008; 5(7):637-43. DOI: 10.1038/nmeth.1227. View

16.

Helman A, Lim B, Andreu M, Kim Y, Shestkin T, Lu H . RTK signaling modulates the Dorsal gradient. Development. 2012; 139(16):3032-9. PMC: 3403108. DOI: 10.1242/dev.075812. View

17.

Kohli V, Robles V, Cancela M, Acker J, Waskiewicz A, Elezzabi A . An alternative method for delivering exogenous material into developing zebrafish embryos. Biotechnol Bioeng. 2007; 98(6):1230-41. DOI: 10.1002/bit.21564. View

18.

Rushlow C, Shvartsman S . Temporal dynamics, spatial range, and transcriptional interpretation of the Dorsal morphogen gradient. Curr Opin Genet Dev. 2012; 22(6):542-6. DOI: 10.1016/j.gde.2012.08.005. View

19.

Chung K, Kim Y, Kanodia J, Gong E, Shvartsman S, Lu H . A microfluidic array for large-scale ordering and orientation of embryos. Nat Methods. 2010; 8(2):171-176. PMC: 4100329. DOI: 10.1038/nmeth.1548. View

20.

Kanodia J, Kim Y, Tomer R, Khan Z, Chung K, Storey J . A computational statistics approach for estimating the spatial range of morphogen gradients. Development. 2011; 138(22):4867-74. PMC: 3201657. DOI: 10.1242/dev.071571. View