3D-printed microwell arrays for Ciona microinjection and timelapse imaging

Area of Science:

• Developmental Biology
• Chordate Embryology
• Marine Biology

Background:

• Ascidians, like Ciona, are vital chordate relatives for studying vertebrate evolution.
• Their small embryos and genomes are advantageous for imaging and genomic analysis.
• Microinjection of Ciona eggs is challenging due to their small size and difficulty in immobilization.

Purpose of the Study:

• To develop a novel device for immobilizing Ciona eggs for microinjection.
• To improve the efficiency and accessibility of functional studies in Ciona.
• To create a tool beneficial for both microinjection and timelapse imaging of multiple embryos.

Main Methods:

• Fabrication of an injection mold using micro-resolution stereolithography.
• Design features include a grid of posts to cast single-egg-sized wells in agarose.
• Testing of the device for its efficacy in restraining eggs during microinjection.

Main Results:

• Successfully developed and tested a 3D-printed agarose mold for Ciona egg microinjection.
• The mold effectively restrains individual eggs, facilitating needle insertion and withdrawal.
• The device also proved useful for timelapse imaging of multiple embryos simultaneously.

Conclusions:

• The developed injection mold significantly enhances the feasibility of microinjection in Ciona.
• This 3D-printed device offers a rapid, inexpensive, and effective solution for Ciona functional studies.
• The mold serves a dual purpose, aiding both microinjection and high-throughput embryo imaging.