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Proceedings Paper

Adaptive optics improves multiphoton super-resolution imaging
Author(s): Wei Zheng; Yicong Wu; Peter Winter; Hari Shroff
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Paper Abstract

Three dimensional (3D) fluorescence microscopy has been essential for biological studies. It allows interrogation of structure and function at spatial scales spanning the macromolecular, cellular, and tissue levels. Critical factors to consider in 3D microscopy include spatial resolution, signal-to-noise (SNR), signal-to-background (SBR), and temporal resolution. Maintaining high quality imaging becomes progressively more difficult at increasing depth (where optical aberrations, induced by inhomogeneities of refractive index in the sample, degrade resolution and SNR), and in thick or densely labeled samples (where out-of-focus background can swamp the valuable, in-focus-signal from each plane). In this report, we introduce our new instrumentation to address these problems. A multiphoton structured illumination microscope was simply modified to integrate an adpative optics system for optical aberrations correction. Firstly, the optical aberrations are determined using direct wavefront sensing with a nonlinear guide star and subsequently corrected using a deformable mirror, restoring super-resolution information. We demonstrate the flexibility of our adaptive optics approach on a variety of semi-transparent samples, including bead phantoms, cultured cells in collagen gels and biological tissues. The performance of our super-resolution microscope is improved in all of these samples, as peak intensity is increased (up to 40-fold) and resolution recovered (up to 176±10 nm laterally and 729±39 nm axially) at depths up to ~250 μm from the coverslip surface.

Paper Details

Date Published: 23 February 2018
PDF: 8 pages
Proc. SPIE 10502, Adaptive Optics and Wavefront Control for Biological Systems IV, 1050211 (23 February 2018); doi: 10.1117/12.2288155
Show Author Affiliations
Wei Zheng, Shenzhen Institutes of Advanced Technology (China)
National Institute of Biomedical Imaging and Bioengineering (United States)
Yicong Wu, National Institute of Biomedical Imaging and Bioengineering (United States)
Peter Winter, National Institute of Biomedical Imaging and Bioengineering (United States)
Hari Shroff, National Institute of Biomedical Imaging and Bioengineering (United States)


Published in SPIE Proceedings Vol. 10502:
Adaptive Optics and Wavefront Control for Biological Systems IV
Thomas G. Bifano; Joel Kubby; Sylvain Gigan, Editor(s)

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