Abstract

Three-dimensional structured illumination microscopy (3D-SIM) is a wide-field super-resolution technique in fluorescent imaging that can double the resolution beyond its classical limit. We introduce, to the best of our knowledge, a new 3D reconstruction algorithm based on the Richardson–Lucy deconvolution. The 3D-SIM imaging principle and the reconstruction steps are demonstrated in detail. Microspheres and biological specimen are used to present the performance of this method. The background of the out-of-focus portion is effectively suppressed, and true optical sectioning and super-resolution can be achieved simultaneously. For the custom-built 3D-SIM and this reconstruction algorithm, the measured resolution was 99.5±5  nm laterally and 294±9  nm axially.

© 2019 Optical Society of America

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Supplementary Material (2)

NameDescription
» Visualization 1       Three-dimensional results of structured illumination microscopy. Each frame represents a Z-plane.
» Visualization 2       Three-dimensional results of wide field microscopy. Each frame represents a Z-plane.

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