STED-like resolution enhancement with focus extension

Kai-Ping Yang; Wei-Kuan Lin; Kuo-Jen Hsu; Yen-Yin Lin; Shi-Wei Chu

doi:10.1117/12.2208865

9 March 2016 STED-like resolution enhancement with focus extension

Kai-Ping Yang, Wei-Kuan Lin, Kuo-Jen Hsu, Yen-Yin Lin, Shi-Wei Chu

Proceedings Volume 9713, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIII; 97131N (2016) https://doi.org/10.1117/12.2208865
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

In recent years, the techniques of super-resolution have generated widespread impacts in science. Stimulated emission depletion (STED) microscopy is known for achieving sub-diffraction-limit resolution by using a donut-shaped beam to deplete the fluorescence around a focal spot while leaving a central part active to emit fluorescence. However, since STED microscopy is based on fluorescence, it suffers from photo-bleaching. We recently developed a new technique and termed it as suppression of scattering imaging (SUSI) microscopy. It uses a STED-like setup and achieves super resolution imaging by utilizing the nonlinearity of scattering from gold nanoparticles. Therefore, SUSI microscopy avoids the photo-bleaching issue. Nonetheless, for fast volumetric imaging, SUSI microscopy is limited with slow axial translation of the objective or sample. Here we combine SUSI microscopy with a refractive-index-variable lens to axially move the focus at very high speed. This combination allows simultaneous observation of tissue dynamics over a three-dimensional volume within one second. The new technique paves the way toward high-speed super-resolution imaging for biological tissues.

Conference Presentation

Citation Download Citation

Kai-Ping Yang, Wei-Kuan Lin, Kuo-Jen Hsu, Yen-Yin Lin, and Shi-Wei Chu "STED-like resolution enhancement with focus extension", Proc. SPIE 9713, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIII, 97131N (9 March 2016); https://doi.org/10.1117/12.2208865

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