Two-photon excitation improves multifocal structured illumination microscopy in thick scattering tissue.
Multifocal structured illumination microscopy (MSIM) provides a twofold resolution enhancement beyond the diffraction limit at sample depths up to 50 µm, but scattered and out-of-focus light in thick samples degrades MSIM performance. Here we implement MSIM with a microlens array to enable efficient two-photon excitation. Two-photon MSIM gives resolution-doubled images with better sectioning and contrast in thick scattering samples such as Caenorhabditis elegans embryos, Drosophila melanogaster larval salivary glands, and mouse liver tissue.
National Institute of Biomedical Imaging and Bioengineering, National Institute of Dental and Craniofacial Research, and National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892.
Maria
M
Ingaramo
Andrew G
AG
York
Peter
P
Wawrzusin
Oleg
O
Milberg
Amy
A
Hong
Roberto
R
Weigert
Hari
H
Shroff
George H
GH
Patterson
D000818:Animals; D017173:Caenorhabditis elegans / Q000196:embryology; D004331:Drosophila melanogaster / Q000254:growth & development; D007814:Larva / Q000737:chemistry; D008029:Lighting; D008099:Liver / Q000737:chemistry; D051379:Mice; D008853:Microscopy / Q000379:methods*; D017785:Photons
multiphoton; superresolution