Microfluidic system with integrated microinjector for automated Drosophila embryo injection

被引:33
作者
Delubac, Daniel [1 ]
Highley, Christopher B. [1 ]
Witzberger-Krajcovic, Melissa [2 ]
Ayoob, Joseph C. [3 ]
Furbee, Emily C. [2 ]
Minden, Jonathan S. [2 ]
Zappe, Stefan [1 ]
机构
[1] Carnegie Mellon Univ, Dept Biomed Engn, Pittsburgh, PA 15213 USA
[2] Carnegie Mellon Univ, Dept Biol Sci, Pittsburgh, PA 15213 USA
[3] Univ Pittsburgh, Dept Computat & Syst Biol, Pittsburgh, PA 15260 USA
基金
美国国家科学基金会;
关键词
RNA; TRANSFORMATION; INTERFERENCE; GENE;
D O I
10.1039/c2lc40104e
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Drosophila is one of the most important model organisms in biology. Knowledge derived from the recently sequenced 12 genomes of various Drosophila species can today be combined with the results of more than 100 years of research to systematically investigate Drosophila biology at the molecular level. In order to enable automated, high-throughput manipulation of Drosophila embryos, we have developed a microfluidic system based on a Pyrex-silicon-Pyrex sandwich structure with integrated, surface-micromachined silicon nitride injector for automated injection of reagents. Our system automatically retrieves embryos from an external reservoir, separates potentially clustered embryos through a sheath flow mechanisms, passively aligns an embryo with the integrated injector through geometric constraints, and pushes the embryo onto the injector through flow drag forces. Automated detection of an embryo at injection position through an external camera triggers injection of reagents and subsequent ejection of the embryo to an external reservoir. Our technology can support automated screens based on Drosophila embryos as well as creation of transgenic Drosophila lines. Apart from Drosophila embryos, the layout of our system can be easily modified to accommodate injection of oocytes, embryos, larvae, or adults of other species and fills an important technological gap with regard to automated manipulation of multicellular organisms.
引用
收藏
页码:4911 / 4919
页数:9
相关论文
共 31 条
[1]   Microfluidic based single cell microinjection [J].
Adamo, Andrea ;
Jensen, Klavs F. .
LAB ON A CHIP, 2008, 8 (08) :1258-1261
[2]  
[Anonymous], 2002, NATURE, V420, P520
[3]   High-throughput RNA interference screens in Drosophila tissue culture cells [J].
Armknecht, S ;
Boutros, M ;
Kiger, A ;
Nybakken, K ;
Mathey-Prevot, B ;
Perrimon, N .
RNA INTERFERENCE, 2005, 392 :55-+
[4]   Site-specific transformation of Drosophila via φC31 integrase-mediated cassette exchange [J].
Bateman, Jack R. ;
Lee, Anne M. ;
Wu, C. -ting .
GENETICS, 2006, 173 (02) :769-777
[5]   Characterization of fluidic microassembly for immobilization and positioning of Drosophila embryos in 2-D arrays [J].
Bernstein, RW ;
Zhang, XJ ;
Zappe, S ;
Fish, M ;
Scott, M ;
Solgaard, O .
SENSORS AND ACTUATORS A-PHYSICAL, 2004, 114 (2-3) :191-196
[6]   Genome-wide RNAi analysis of growth and viability in Drosophila cells [J].
Boutros, M ;
Kiger, AA ;
Armknecht, S ;
Kerr, K ;
Hild, M ;
Koch, B ;
Haas, SA ;
Paro, R ;
Perrimon, N .
SCIENCE, 2004, 303 (5659) :832-835
[7]   ZFIN: enhancements and updates to the zebrafish model organism database [J].
Bradford, Yvonne ;
Conlin, Tom ;
Dunn, Nathan ;
Fashena, David ;
Frazer, Ken ;
Howe, Douglas G. ;
Knight, Jonathan ;
Mani, Prita ;
Martin, Ryan ;
Moxon, Sierra A. T. ;
Paddock, Holly ;
Pich, Christian ;
Ramachandran, Sridhar ;
Ruef, Barbara J. ;
Ruzicka, Leyla ;
Schaper, Holle Bauer ;
Schaper, Kevin ;
Shao, Xiang ;
Singer, Amy ;
Sprague, Judy ;
Sprunger, Brock ;
Van Slyke, Ceri ;
Westerfield, Monte .
NUCLEIC ACIDS RESEARCH, 2011, 39 :D822-D829
[8]   Genome sequence of the nematode C-elegans:: A platform for investigating biology [J].
不详 .
SCIENCE, 1998, 282 (5396) :2012-2018
[9]   Systematic genome-wide screens of gene function [J].
Carpenter, AE ;
Sabatini, DM .
NATURE REVIEWS GENETICS, 2004, 5 (01) :11-22
[10]   Design and operation of a microfluidic sorter for Drosophila embryos [J].
Chen, CC ;
Zappe, S ;
Sahin, O ;
Zhang, XJ ;
Fish, M ;
Scott, M ;
Solgaard, O .
SENSORS AND ACTUATORS B-CHEMICAL, 2004, 102 (01) :59-66