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Light-sheet microscopy for everyone? Experience of building an OpenSPIM to study flatworm development
Johannes Girstmair1, Anne Zakrzewski1, François Lapraz1,2
1Department of Genetics, Evolution and Environment, University College London, London, WC1E 6BT, UK.
BMC Developmental Biology
|July 2, 2016
Summary
Building an OpenSPIM microscope offers accessible light-sheet microscopy for researchers. This customizable, open-access platform enables high-resolution 3D and 4D imaging for developmental studies, despite some construction challenges.
Area of Science:
- Biotechnology
- Microscopy
- Developmental Biology
Background:
- Selective plane illumination microscopy (SPIM) reduces phototoxicity for extended live imaging.
- OpenSPIM provides an open-access platform for building customizable SPIM microscopes.
- This study details the design, construction, and application of an OpenSPIM for polyclad flatworm research.
Purpose of the Study:
- To describe the process of building and configuring an OpenSPIM microscope.
- To demonstrate the capabilities of a customized OpenSPIM for high-resolution 3D and 4D imaging.
- To investigate the early development of the polyclad flatworm Maritigrella crozieri.
Main Methods:
- Construction of a modified OpenSPIM with dual-sided, two-color laser illumination.
- High-speed multi-view time-lapse imaging of Maritigrella crozieri embryos.
- 3D and 4D reconstruction using open-source software (Fiji).
Main Results:
- The customized OpenSPIM achieved high-resolution 3D and 4D imaging.
- Dual-sided, two-color illumination enhanced signal intensity and detection.
- Stereotypical spiral cleavage in M. crozieri embryos was successfully captured and reconstructed.
Conclusions:
- OpenSPIM and open-source software make light-sheet microscopy accessible.
- Building a custom SPIM is a significant but worthwhile investment.
- Multi-view 4D microscopy presents challenges but is achievable with experience.
