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Imaging adult C. elegans live using light-sheet microscopy
J VAN Krugten1, K-K H Taris1, Erwin J G Peterman1
1LaserLaB and Department of Physics and Astronomy, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands.
Journal of Microscopy
|September 19, 2020
Summary
We developed a straightforward light-sheet microscope for imaging living C. elegans, offering enhanced signal-to-background ratio and reduced phototoxicity compared to traditional methods. This new instrument enables fast, volumetric imaging of neuronal structures in nematodes.
Area of Science:
- Biophysics
- Microscopy
- Developmental Biology
Background:
- Live observation of biological phenomena is crucial for understanding complex physiological mechanisms.
- Traditional fluorescence microscopy techniques like confocal and widefield suffer from low signal-to-background ratio (SBR), phototoxicity, and bleaching due to out-of-focus illumination.
- Light-sheet microscopy (LSM) offers an alternative by illuminating only the focal plane, improving SBR and reducing photobleaching.
Purpose of the Study:
- To present a straightforward light-sheet microscope with a 1.0-NA detection objective and a four-degrees-of-freedom sample-positioning stage.
- To demonstrate the instrument's capability for fast, volumetric imaging of larger living biological samples.
- To evaluate the instrument's utility for imaging the sensory cilia and nervous system of living adult C. elegans.
Main Methods:
- Construction of a light-sheet fluorescence microscope based on the OpenSPIM initiative.
- Integration of a 1.0-NA detection objective and a fast sample-positioning stage with four degrees of freedom.
- Imaging of neuronal structures and sensory cilia in living young adult C. elegans.
Main Results:
- The developed light-sheet microscope achieved high signal-to-background ratio and minimized photobleaching.
- The instrument enabled fast, volumetric imaging of C. elegans neuronal cell bodies and other larger cellular structures.
- Comparison with standard epifluorescence microscopy showed the LSM's superior performance for imaging dynamic biological processes.
Conclusions:
- The presented light-sheet microscope is well-suited for high-resolution, three-dimensional imaging of living C. elegans over extended periods.
- This instrument provides a valuable addition to existing fluorescence microscopy techniques for studying complex biological systems.
- The straightforward design and enhanced imaging capabilities make it a powerful tool for live biological observations.
