Improved 3D cellular morphometry of Caenorhabditis elegans embryos using a refractive index matching medium
Rain Xiong1,2, Kenji Sugioka1,2
1Life Sciences Institute, The University of British Columbia, Vancouver, BC, Canada.
Plos One
|September 30, 2020
Summary
We developed a 3D live imaging method for Caenorhabditis elegans using a refractive index matching medium (RIMM). This technique improves cellular morphometry and reveals high cortical tension in germ cell precursors during development.
Area of Science:
- Developmental Biology
- Cell Biology
- Biophysics
Background:
- Cell shape change is crucial for animal morphogenesis.
- Caenorhabditis elegans is a key model organism for studying developmental mechanisms.
- Standard 3D live imaging of C. elegans is limited by spherical aberrations, hindering cellular morphometry.
Purpose of the Study:
- To develop an improved 3D live imaging method for C. elegans embryos.
- To minimize spherical aberrations caused by refractive index mismatch.
- To enable high-resolution cellular morphometry in 3D for C. elegans.
Main Methods:
- Determined the optimal composition of a refractive index matching medium (RIMM).
- Implemented 3D live imaging of C. elegans embryos using the RIMM.
- Performed 3D cellular morphometry and 2D cell shape simulations.
Main Results:
- The RIMM significantly reduced spherical aberrations and improved signal intensity in deeper cell layers.
- 3D cell segmentation quality was enhanced using images acquired with the RIMM.
- Analysis revealed exceptionally high cortical tension in the germ cell precursor P4.
Conclusions:
- The RIMM is a cost-effective solution for enhancing 3D cellular morphometry in C. elegans.
- This method facilitates a deeper understanding of C. elegans morphogenesis through cell shape analysis.
- The findings provide new insights into the biophysical properties of embryonic cells.
