Low-cost Polyethylene Terephthalate Lamination Microfluidics Designs for Multiplexed Zebrafish Imaging

Area of Science:

• Development of novel research tools for developmental biology and immunology.

Background:

• Zebrafish embryos are ideal for noninvasive intravital imaging due to their transparency.
• Microfluidic devices facilitate long-term imaging of zebrafish but traditional fabrication is complex and costly.
• Accessible fabrication methods are needed to broaden the use of these imaging techniques.

Purpose of the Study:

• To develop a low-cost, accessible microfluidic device for long-term zebrafish embryo imaging.
• To enable detailed observation of fundamental biological processes like wound healing and immune cell migration.

Main Methods:

• Adapted a low-cost polyethylene terephthalate lamination method for microfluidic device fabrication.
• Designed the Rotational Assistant for Danio Imaging of Subsequent Healing (RADISH) device.
• Utilized the RADISH device for drug treatment, manual wounding, and imaging of up to four zebrafish embryos simultaneously.

Main Results:

• Successfully captured calcium signaling dynamics around laser ablation and transection wounds within 2 hours post-injury.
• Monitored neutrophil recruitment to wound sites for up to 24 hours.
• Demonstrated the device's capability for long-term, multi-embryo imaging and experimental manipulation.

Conclusions:

• The RADISH device offers an accessible and cost-effective alternative for microfluidic fabrication.
• This method significantly lowers the technical barrier for intravital imaging of zebrafish embryos.
• Facilitates advanced research into wound healing and immune responses in a developmental context.