120nm resolution in thick samples with structured illumination and adaptive optics

Benjamin Thomas; Megan Sloan; Adrian J. Wolstenholme; Peter Kner

doi:10.1117/12.2040931

7 March 2014 120nm resolution in thick samples with structured illumination and adaptive optics

Benjamin Thomas, Megan Sloan, Adrian J. Wolstenholme, Peter Kner

Proceedings Volume 8978, MEMS Adaptive Optics VIII; 89780H (2014) https://doi.org/10.1117/12.2040931
Event: SPIE MOEMS-MEMS, 2014, San Francisco, California, United States

Abstract

μLinear Structured Illumination Microscopy (SIM) provides a two-fold increase over the diffraction limited resolution. SIM produces excellent images with 120nm resolution in tissue culture cells in two and three dimensions. For SIM to work correctly, the point spread function (PSF) and optical transfer function (OTF) must be known, and, ideally, should be unaberrated. When imaging through thick samples, aberrations will be introduced into the optical system which will reduce the peak intensity and increase the width of the PSF. This will lead to reduced resolution and artifacts in SIM images. Adaptive optics can be used to correct the optical wavefront restoring the PSF to its unaberrated state, and AO has been used in several types of fluorescence microscopy. We demonstrate that AO can be used with SIM to achieve 120nm resolution through 25m of tissue by imaging through the full thickness of an adult C. elegans roundworm. The aberrations can be corrected over a 25μm × 45μm field of view with one wavefront correction setting, demonstrating that AO can be used effectively with widefield superresolution techniques.

Citation Download Citation

Benjamin Thomas, Megan Sloan, Adrian J. Wolstenholme, and Peter Kner "120nm resolution in thick samples with structured illumination and adaptive optics", Proc. SPIE 8978, MEMS Adaptive Optics VIII, 89780H (7 March 2014); https://doi.org/10.1117/12.2040931

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