Automating fruit fly Drosophila embryo injection for high throughput transgenic studies.

نویسندگان

  • E Cornell
  • W W Fisher
  • R Nordmeyer
  • D Yegian
  • M Dong
  • M D Biggin
  • S E Celniker
  • J Jin
چکیده

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.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Concentration dependent effect of morphine, aspirin, capsaicin and chili pepper hydro alcoholic extract on thermal and chemical pain model in fruit fly (Drosophila melanogaster)

Introduction: Pain research using animal models is related to ethical concerns, so invertebrates and insects have been recommended by researchers. In the present study, the nociceptive and antinociceptive effects of capsaicin, aspirin, morphine and chili extract were examined using fruit fly (Drosophila melanogaster) as an alternative for rodent pain model. Methods: Stage 3 of larvae and ad...

متن کامل

FLIC: High-Throughput, Continuous Analysis of Feeding Behaviors in Drosophila

We present a complete hardware and software system for collecting and quantifying continuous measures of feeding behaviors in the fruit fly, Drosophila melanogaster. The FLIC (Fly Liquid-Food Interaction Counter) detects analog electronic signals as brief as 50 µs that occur when a fly makes physical contact with liquid food. Signal characteristics effectively distinguish between different type...

متن کامل

Preservation of plasma membrane ultrastructure in Drosophila embryos and larvae prepared by high - pressure freezing and freeze - substitution

The ultrastructural analysis of biological structures is often critical to the understanding of biological processes. Fixation by freeze-substitution after immobilisation by High-pressure freezing (HPF-FS) is a powerful method to preserve the ultrastructure of small specimens such as the embryo of the fruit fly Drosophila melanogaster. But only a small fraction of specimens prepared following t...

متن کامل

Imaging of Transgenic Cricket Embryos Reveals Cell Movements Consistent with a Syncytial Patterning Mechanism

The mode of insect embryogenesis varies among species, reflecting adaptations to different life history strategies [1, 2]. In holometabolous insects, which include the model systems, such as the fruit fly and the red flour beetle, a large proportion of the blastoderm produces an embryo, whereas hemimetabolous embryos generally arise from a small region of the blastoderm [3]. Despite their impor...

متن کامل

A " Model " Model System to Study Neurodegeneration

T he fruit fly, Drosophila melanogaster, is a powerful model genetic organism that has been used since the turn of the previous century in the study of complex biological problems. In the last decade, numerous researchers have focused their attention on understanding neurodegenerative diseases by utilizing this model system. Numerous Drosophila mutants have been isolated that profoundly affect ...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • The Review of scientific instruments

دوره 79 1  شماره 

صفحات  -

تاریخ انتشار 2008