Automating fruit fly Drosophila embryo injection for high throughput transgenic studies.
To decipher and manipulate the 14 000 identified Drosophila genes, there is a need to inject a large number of embryos with transgenes. We have developed an automated instrument for high throughput injection of Drosophila embryos. It was built on an inverted microscope, equipped with a motorized xy stage, autofocus, a charge coupled device camera, and an injection needle mounted on a high speed vertical stage. A novel, micromachined embryo alignment device was developed to facilitate the arrangement of a large number of eggs. The control system included intelligent and dynamic imaging and analysis software and an embryo injection algorithm imitating a human operator. Once the injection needle and embryo slide are loaded, the software automatically images and characterizes each embryo and subsequently injects DNA into all suitable embryos. The ability to program needle flushing and monitor needle status after each injection ensures reliable delivery of biomaterials. Using this instrument, we performed a set of transformation injection experiments. The robot achieved injection speeds and transformation efficiencies comparable to those of a skilled human injector. Because it can be programed to allow injection at various locations in the embryo, such as the anterior pole or along the dorsal or ventral axes, this system is also suitable for injection of general biochemicals, including drugs and RNAi.
Cornell
E
E
Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720, USA.
Fisher
W W
WW
Nordmeyer
R
R
Yegian
D
D
Dong
M
M
Biggin
M D
MD
Celniker
S E
SE
Jin
J
J
D000818:Animals; D030801:Animals, Genetically Modified; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics; D004330:Drosophila / Q000196:embryology* / Q000235:genetics*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007267:Injections / Q000295:instrumentation* / Q000379:methods; D015203:Reproducibility of Results; D012371:Robotics / Q000295:instrumentation* / Q000379:methods; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000295:instrumentation / Q000379:methods; D014162:Transfection / Q000295:instrumentation* / Q000379:methods
D004247:DNA