Stimulated emission depletion microscopy with a supercontinuum source and fluorescence lifetime imaging

Abstract

We demonstrate stimulated emission depletion (STED) microscopy implemented in a laser scanning confocal microscope using excitation light derived from supercontinuum generation in a microstructured optical fiber. Images with resolution improvement beyond the far-field diffraction limit in both the lateral and axial directions were acquired by scanning overlapped excitation and depletion beams in two dimensions using the flying spot scanner of a commercially available laser scanning confocal microscope. The spatial properties of the depletion beam were controlled holographically using a programmable spatial light modulator, which can rapidly change between different STED imaging modes and also compensate for aberrations in the optical path. STED fluorescence lifetime imaging microscopy is demonstrated through the use of time-correlated single photon counting.

© 2008 Optical Society of America

Evaluating image resolution in stimulated emission depletion microscopy

Giorgio Tortarolo, Marco Castello, Alberto Diaspro, Sami Koho, and Giuseppe Vicidomini
Optica 5(1) 32-35 (2018)

Fast, super resolution imaging via Bessel-beam stimulated emission depletion microscopy

P. Zhang, P. M. Goodwin, and J. H. Werner
Opt. Express 22(10) 12398-12409 (2014)

Supercontinuum source tuned by an on-axis monochromator for fluorescence lifetime imaging

Raffaella Mercatelli, Silvia Soria, Giuseppe Molesini, Federica Bianco, Giancarlo Righini, and Franco Quercioli
Opt. Express 18(19) 20505-20511 (2010)

STED microscopy with a supercontinuum laser source

Dominik Wildanger, Eva Rittweger, Lars Kastrup, and Stefan W. Hell
Opt. Express 16(13) 9614-9621 (2008)

Dual-mode super-resolution imaging with stimulated emission depletion microscopy and fluorescence emission difference microscopy

Yifan Wang, Ye Ma, Cuifang Kuang, Yue Fang, Yingke Xu, Xu Liu, and Zhihua Ding
Appl. Opt. 54(17) 5425-5431 (2015)