Adaptive optics improves multiphoton super-resolution imaging

• Published in: Nature methods Vol. 14; no. 9; pp. 869 - 872
• Main Authors: Zheng, Wei, Wu, Yicong, Winter, Peter, Fischer, Robert, Nogare, Damian Dalle, Hong, Amy, McCormick, Chad, Christensen, Ryan, Dempsey, William P, Arnold, Don B, Zimmerberg, Joshua, Chitnis, Ajay, Sellers, James, Waterman, Clare, Shroff, Hari
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01-09-2017; Nature Publishing Group
• Subjects: 14/69; 631/1647/245/2226; 631/1647/328/2238; Adaptive optics; Bioengineering; Bioinformatics; Biological Microscopy; Biological Techniques; Biomedical Engineering/Biotechnology; Brain; brief-communication; Collagen; Deformation; Embryos; Equipment Design; Equipment Failure Analysis; Feedback; Formability; Fruit flies; Gels; Image Enhancement - instrumentation; Image Enhancement - methods; Image resolution; Insects; Laboratories; Larvae; Lenses; Life Sciences; Methods; Microscopy; Microscopy, Fluorescence, Multiphoton - instrumentation; Microscopy, Fluorescence, Multiphoton - methods; Neuroimaging; Optics; Photons; Properties; Proteomics; Reproducibility of Results; Sensitivity and Specificity; Zebrafish; United States
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/nmeth.4337

Adaptive optics and two-photon instant structured illumination microscopy are combined to provide improved super-resolution imaging within optically aberrating biological samples.
We improve multiphoton structured illumination microscopy using a nonlinear guide star to determine optical aberrations and a deformable mirror to correct them. We demonstrate our method on bead phantoms, cells in collagen gels, nematode larvae and embryos,
Drosophila
brain, and zebrafish embryos. Peak intensity is increased (up to 40-fold) and resolution recovered (up to 176 ± 10 nm laterally, 729 ± 39 nm axially) at depths ∼250 μm from the coverslip surface.