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Video-rate Scanning Confocal Microscopy and Microendoscopy
Published on: October 20, 2011
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Video-rate remote refocusing through continuous oscillation of a membrane deformable mirror
Terry Wright1, Hugh Sparks1, Carl Paterson1
1Photonics Group, Department of Physics, Imperial College London, South Kensington Campus, London SW7 2AZ, United Kingdom.
Jphys Photonics
|October 25, 2021
Summary
This study demonstrates rapid microscope refocusing using a deformable mirror (DM), achieving high-quality imaging over a wide defocus range. This technology could enable faster 3D imaging in advanced microscopy techniques.
Area of Science:
- Optical microscopy
- Biomedical imaging
Background:
- Microscopy requires precise focusing, especially with high numerical aperture (NA) objectives.
- Rapid refocusing is crucial for advanced imaging techniques like light-sheet fluorescence microscopy (LSFM).
Purpose of the Study:
- To present a deformable mirror (DM) system for rapid microscope refocusing.
- To evaluate the performance of a membrane DM for high-NA objectives.
- To demonstrate the potential for video-rate volumetric imaging.
Main Methods:
- Utilized an Alpao DM97-15 membrane DM with a 40x/0.80 NA water-immersion objective.
- Achieved refocusing over a -50 to 50 micrometer range at 26.3 sweeps/second.
- Implemented continuous mirror sweeping to mitigate hysteresis issues.
Main Results:
- Maintained a mean Strehl metric >0.6 over a 200x200 micrometer field of view.
- Demonstrated effective refocusing across a 77 micrometer defocus range.
- Successfully addressed membrane DM hysteresis through an optimization procedure.
Conclusions:
- Deformable mirror-based refocusing is effective for high-NA microscopy.
- The system enables rapid, high-quality imaging over extended defocus ranges.
- This technology holds promise for advancing volumetric imaging in LSFM.
