Journal
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 83, Pages -Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/50841
Keywords
Developmental Biology; Issue 83; Sea Urchins; microinjection; sea urchin embryos; treatment delivery; high throughput; mouth pipette; DNA constructs; mRNAs; morpholino antisense oligonucleotides
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Funding
- University of Delaware Research Fund
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Microinjection into cells and embryos is a common technique that is used to study a wide range of biological processes. In this method a small amount of treatment solution is loaded into a microinjection needle that is used to physically inject individual immobilized cells or embryos. Despite the need for initial training to perform this procedure for high-throughput delivery, microinjection offers maximum efficiency and reproducible delivery of a wide variety of treatment solutions (including complex mixtures of samples) into cells, eggs or embryos. Applications to microinjections include delivery of DNA constructs, mRNAs, recombinant proteins, gain of function, and loss of function reagents. Fluorescent or colorimetric dye is added to the injected solution to enable instant visualization of efficient delivery as well as a tool for reliable normalization of the amount of the delivered solution. The described method enables microinjection of 100-400 sea urchin zygotes within 10-15 min.
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