3-D stimulated emission depletion microscopy with programmable aberration correction

• Published in: Journal of biophotonics Vol. 7; no. 1-2; pp. 29 - 36
• Main Authors: 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.
• Format: Journal Article
• Language: English
• Published: Berlin WILEY-VCH Verlag 01.01.2014; WILEY‐VCH Verlag; Wiley Subscription Services, Inc
• Subjects: Cell Line; confocal microscopy; fluorescence microscopy; Image Processing, Computer-Assisted - methods; Imaging, Three-Dimensional - methods; immune synapse; Killer Cells, Natural - cytology; Microscopy - methods; Microspheres; Nanodiamonds - chemistry; nanoscopy; STED microscopy; super-resolution; nanoscopy; STED microscopy; confocal microscopy; fluorescence microscopy; immune synapse; super-resolution
• ISSN: 1864-063X; 1864-0648
• DOI: 10.1002/jbio.201300041

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)