Broad applicability of a streamlined ethyl cinnamate-based clearing procedure

Area of Science:

• Biomedical Imaging
• Molecular Biology
• Developmental Biology

Background:

• Tissue turbidity and opacity limit deep microscopic imaging.
• Existing tissue clearing methods can be costly, time-consuming, and complex.
• Current methods may negatively impact fluorescence signal preservation.

Purpose of the Study:

• To develop an accessible and efficient tissue clearing method.
• To overcome limitations of existing tissue clearing techniques.
• To enable high-resolution 3D imaging of large biological samples.

Main Methods:

• Developed 2Eci (2nd generation ethyl cinnamate-based clearing).
• Applied 2Eci to various species including human organoids, Drosophila melanogaster, zebrafish, axolotl, and Xenopus laevis.
• Evaluated preservation of fluorescent proteins like GFP, mCherry, Brainbow, and Alexa-conjugated fluorophores.

Main Results:

• 2Eci cleared diverse tissues from multiple species in 1-5 days.
• Broad-spectrum fluorescence preservation was achieved, including GFP, mCherry, Brainbow, and Alexa fluorophores.
• Ethyl cinnamate was found to be non-toxic and suitable for multi-user facilities.

Conclusions:

• 2Eci offers a rapid, cost-effective, and user-friendly tissue clearing solution.
• The method is broadly applicable across various species and fluorescent labels.
• 2Eci significantly expands accessibility to advanced 3D tissue imaging for researchers.