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We describe the design and implementation of a stimulated emission depletion (STED) microscope which allows simultaneous three-dimensional super-resolution imaging in two colors. A super-continuum laser source is used to provide all spectral bands necessary for excitation and efficient depletion to achieve a lateral and axial resolution of ~35 nm and ~90 nm respectively. We characterize the systems' performance by imaging colloidal particles and single fluorescent molecules. Its biological applicability is demonstrated by dual-color imaging of nuclear pore complexes and of DNA replication sites in mammalian cells.

Type

Journal article

Journal

Opt Express

Publication Date

24/03/2014

Volume

22

Pages

7028 - 7039

Keywords

Animals, Cell Line, Tumor, Cell Nucleus, Color, DNA, DNA Replication, Humans, Imaging, Three-Dimensional, Mice, Microscopy, S Phase