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J Biophotonics. 2018 Nov;11(11):e201800087. doi: 10.1002/jbio.201800087. Epub 2018 Jul 31.

easySLM-STED: Stimulated emission depletion microscopy with aberration correction, extended field of view and multiple beam scanning.

Author information

1
Photonics Group, Department of Physics, Imperial College London, London, UK.
2
Femtosecond Optics Group, Department of Physics, Imperial College London, London, UK.
3
London Institute of Medical Sciences, Imperial College London, London, UK.
4
Centre for Pathology, Imperial College London, London, UK.

Abstract

We demonstrate a simplified set-up for STED microscopy with a straightforward alignment procedure that uses a single spatial light modulator (SLM) with collinear incident excitation and depletion beams to provide phase modulation of the beam profiles and correction of optical aberrations. We show that this approach can be used to extend the field of view for STED microscopy by correcting chromatic aberration that otherwise leads to walk-off between the focused excitation and depletion beams. We further show how this arrangement can be adapted to increase the imaging speed through multibeam excitation and depletion. Fine adjustments to the alignment can be accomplished using the SLM only, conferring the potential for automation.

KEYWORDS:

STED; aberration correction; spatial light modulator; super resolved microscopy

PMID:
29978591
DOI:
10.1002/jbio.201800087
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