Comprehensive 3D Imaging of Whole Zebrafish Using a Water-Based Clearing Reagent for Hard Tissues.

Area of Science:

• Developmental Biology
• Biomedical Imaging
• Zebrafish Model Organisms

Background:

• Zebrafish (Danio rerio) are crucial for studying development due to transparent embryos.
• Opaque tissues in later zebrafish development hinder deep tissue visualization.
• Existing tissue clearing methods struggle with dense tissues and can cause signal loss.

Purpose of the Study:

• To introduce a novel imaging technique for high-resolution 3D visualization of zebrafish.
• To evaluate the efficacy of the LUCID clearing reagent for dense tissues and fluorescent signal preservation.

Main Methods:

• Utilized a new clearing reagent, LUCID, for larval and juvenile zebrafish.
• Assessed LUCID's ability to clear dense tissues like bone and cartilage.
• Evaluated signal integrity for fluorescent proteins, DNA/actin dyes, and FISH.

Main Results:

• LUCID successfully cleared dense tissues, including pharyngeal cartilage and tooth bone.
• The method preserved morphological integrity without significant distortion.
• No significant loss of fluorescent signal was observed across various labeling methods.

Conclusions:

• LUCID offers a powerful new tool for complete cellular-resolution 3D imaging in zebrafish.
• This technique overcomes limitations of previous clearing methods, enabling deeper insights into zebrafish internal structures.
• LUCID facilitates high-resolution imaging of whole, fluorescently stained zebrafish, advancing developmental research.