Effects of donor and acceptor's fluorescence lifetimes on the method of applying Förster resonance energy transfer in STED microscopy

Suhui Deng; Jianfang Chen; Zhaoshuai Gao; Chunhai Fan; Qiurong Yan; Yuhao Wang

doi:10.1111/jmi.12608

Effects of donor and acceptor's fluorescence lifetimes on the method of applying Förster resonance energy transfer in STED microscopy

J Microsc. 2018 Jan;269(1):59-65. doi: 10.1111/jmi.12608. Epub 2017 Jul 31.

Authors

Suhui Deng¹, Jianfang Chen², Zhaoshuai Gao³, Chunhai Fan³, Qiurong Yan¹, Yuhao Wang¹

Affiliations

¹ School of Information Engineering, Nanchang University, Nanchang, China.
² Chinese Academy of Sciences, Shanghai Institute of Optics and Fine Mechanics, Shanghai, China.
³ Division of Physical Biology, and Bioimaging Center, Shanghai Synchrotron Radiation Facility, Chinese Academy of Sciences, Shanghai Institute of Applied Physics, Shanghai, China.

PMID: 28758683
DOI: 10.1111/jmi.12608

Abstract

Förster resonance energy transfer (FRET) probes being used to improve the resolution of stimulated emission depletion (STED) microscopy are numerically discussed. Besides the FRET efficiency and the excitation intensity, the fluorescence lifetimes of donor and acceptor are found to be another key parameter for the resolution enhancement. Using samples of FRET pairs with shorter donor lifetime and longer acceptor lifetime enhances the nonlinearity of the donor fluorescence, which leads to an increased resolution. The numerical simulation shows that a double resolution improvement of STED microscopy can be achieved by using Cy3-Atto647N samples when compared with that of using standard Cy3-only samples.

Keywords: Fluorescence lifetime; Förster resonance energy transfer (FRET); nonlinear donor fluorescence; stimulated emission depletion (STED) microscopy; superresolution.

Publication types

Research Support, Non-U.S. Gov't