Open AccessStable stimulated emission depletion imaging of extended sample regions
Author(s) -
Jonatan Alvelid,
Ilaria Testa
Publication year - 2019
Publication title -
journal of physics. d, applied physics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.857
H-Index - 198
eISSN - 1361-6463
pISSN - 0022-3727
DOI - 10.1088/1361-6463/ab4c13
Subject(s) - sted microscopy , sample (material) , image resolution , optics , focus (optics) , cardinal point , resolution (logic) , stimulated emission , plane (geometry) , field of view , physics , computer science , artificial intelligence , laser , mathematics , geometry , thermodynamics
Stimulated emission depletion (STED) nanoscopy has become one of the most used nanoscopy techniques over the last decade. However, most recordings are done in specimen regions no larger than 10–30 × 10–30 μ m 2 due to aberrations, instability and manual mechanical stages. Here, we demonstrate automated 2D and 3D STED nanoscopy of extended sample regions up to 0.5 × 0.5 mm 2 by using a scanning system that maintains stationary beams in the back focal plane. The setup allows up to 80–100 × 80–100 μ m 2 field of view (FOV) with uniform spatial resolution, a mechanical stage allowing sequential tiling to record larger sample areas, and a feedback system keeping the sample in focus at all times. Taken together, this allows automated recording of theoretically unlimited-sized sample areas and volumes, without compromising the achievable spatial resolution and image quality.