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Adaptive optics in an oblique plane microscope
Conor Mcfadden1,2, Zach Marin1,3, Bingying Chen1,2
1Lyda Hill Department for Bioinformatics, UT Southwestern Medical Center, 6000 Harry Hines BLVD, Dallas, TX 75390, USA.
Biomedical Optics Express
|September 30, 2024
Summary
Adaptive optics (AO) corrects aberrations in oblique plane microscopy (OPM), a 3D light-sheet fluorescence microscopy technique. This simplifies the system and improves imaging of biological samples like zebrafish embryos.
Area of Science:
- Biomedical Optics
- Microscopy
- Optical Engineering
Background:
- Adaptive optics (AO) enhances imaging resolution in microscopy by correcting optical aberrations.
- Light-sheet fluorescence microscopy (LSFM) is a 3D imaging technique, but aberrations in separate illumination and detection paths complicate performance.
- Oblique plane microscopy (OPM) offers a simplified LSFM approach with a single objective lens.
Purpose of the Study:
- To investigate the application of adaptive optics (AO) in oblique plane microscopy (OPM) for improved 3D imaging.
- To demonstrate a simplified AO system that corrects aberrations in both illumination and detection paths simultaneously.
- To assess the impact of AO-corrected OPM on imaging resolution and signal strength in biological samples.
Main Methods:
- Implemented a single deformable mirror for simultaneous aberration correction in both illumination and detection paths of an OPM system.
- Utilized a projection imaging mode to stabilize and enhance wavefront correction in a sensorless AO configuration.
- Tested the AO-OPM system on fluorescent nanospheres and performed live imaging of zebrafish embryos.
Main Results:
- AO correction in OPM restored diffraction-limited resolution for imaging beyond superficial layers.
- A single deformable mirror successfully corrected aberrations in both illumination and detection paths, simplifying the microscope setup.
- The AO-OPM system improved signal strength twofold and enhanced the alignment between the light-sheet and focal plane.
- Imaging of zebrafish vasculature and cancer cells demonstrated the system's capability for advanced biological research.
Conclusions:
- Adaptive optics significantly enhances the performance of oblique plane microscopy by simplifying aberration correction.
- AO in OPM restores optical quality and improves signal detection, enabling deeper and clearer 3D imaging of biological specimens.
- This integrated AO-OPM approach provides a powerful tool for advanced 3D fluorescence microscopy in biological and biomedical research.
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