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3‐D stimulated emission depletion microscopy with programmable aberration correction
Author(s) -
Lenz Martin O.,
Sinclair Hugo G.,
Savell Alexander,
Clegg James H.,
Brown Alice C. N.,
Davis Daniel M.,
Dunsby Chris,
Neil Mark A. A.,
French Paul M. W.
Publication year - 2014
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201300041
Subject(s) - sted microscopy , microscopy , optics , super resolution microscopy , microscope , stimulated emission , resolution (logic) , point spread function , materials science , optical microscope , fluorescence microscope , image resolution , phase (matter) , immunological synapse , chemistry , fluorescence , physics , scanning confocal electron microscopy , scanning electron microscope , biology , laser , organic chemistry , artificial intelligence , computer science , t cell receptor , immune system , t cell , immunology
We present a stimulated emission depletion (STED) microscope that provides 3‐D super resolution by simultaneous depletion using beams with both a helical phase profile for enhanced lateral resolution and an annular phase profile to enhance axial resolution. The 3‐D depletion point spread function is realised using a single spatial light modulator that can also be programmed to compensate for aberrations in the microscope and the sample. We apply it to demonstrate the first 3‐D super‐resolved imaging of an immunological synapse between a Natural Killer cell and its target cell. (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)