Two-photon excitation improves multifocal structured illumination microscopy in thick scattering tissue.

Ingaramo, Maria; York, Andrew G; Wawrzusin, Peter; Milberg, Oleg; Hong, Amy; Weigert, Roberto; Shroff, Hari; Patterson, George H · Proc Natl Acad Sci U S A · 2014

basic_science · Level V

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Abstract

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.

Medical subject headings