Far-field optical nanoscopy based on continuous wave laser stimulated emission depletion.
Stimulated emission depletion (STED) microscopy is one of the breakthrough technologies that belong to far-field optical microscopy and can achieve nanoscale spatial resolution. We demonstrate a far-field optical nanoscopy based on continuous wave lasers with different wavelengths, i.e., violet and green lasers for excitation and STED, respectively. Fluorescent dyes Coumarin 102 and Atto 390 are used for validating the depletion efficiency. Fluorescent nanoparticles are selected for characterizing the spatial resolution of the STED system. Linear scanning of the laser beams of the STED system along one line of a microscope slide, which is coated with the nanoparticles, indicates that a spatial resolution of about 70 nm has so far been achieved. A two-dimensional image of the particle pattern of the STED system is constructed and compared with scanning confocal microscope. The present work has further extended the application of the STED microscopy into the blue regime.
Department of Mechanical Engineering and Biomedical Engineering Program, University of South Carolina, Columbia, South Carolina 29208, USA.
Cuifang
C
Kuang
Wei
W
Zhao
Guiren
G
Wang
D004867:Equipment Design; D019544:Equipment Failure Analysis; D007089:Image Enhancement / Q000295:instrumentation*; D008029:Lighting / Q000295:instrumentation*; D018613:Microscopy, Confocal / Q000295:instrumentation*; D008856:Microscopy, Fluorescence / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity