在商业双光子 FLIM 显微镜上进行可行的附加升级，以实现 CFP-YFP 对的最佳 FLIM-FRET 成像。

A feasible add-on upgrade on a commercial two-photon FLIM microscope for optimal FLIM-FRET imaging of CFP-YFP pairs.

机构信息

Britton Chance Center for Biomedical Photonics, Wuhan National Laboratory for Optoelectronics-Huazhong University of Science and Technology, Wuhan, China.

出版信息

J Fluoresc. 2013 May;23(3):543-9. doi: 10.1007/s10895-013-1188-8. Epub 2013 Mar 3.

DOI:10.1007/s10895-013-1188-8

PMID:23456419

Abstract

Fluorescence lifetime imaging microscopy (FLIM) based on time-correlated single photon counting (TCSPC) is a widely used method for fluorescence resonance energy transfer (FRET). Here we report a feasible add-on approach to upgrade a commercial two-photon FLIM microscope into a single-photon FLIM microscope which provides optimal FLIM-FRET imaging of FRET pairs consisting of cyan fluorescent proteins (CFPs) as the donor and yellow fluorescent proteins (YFPs) as the acceptor. The capability of the upgraded system is evaluated and discussed, and the imaging performance of the system is demonstrated using FLIM-FRET experiments with a representative CFP-YFP FRET pair (mCerulean-mCitrine).

摘要

基于时间相关单光子计数（TCSPC）的荧光寿命成像显微镜（FLIM）是一种广泛用于荧光共振能量转移（FRET）的方法。在这里，我们报告了一种可行的附加方法，可将商业双光子 FLIM 显微镜升级为单光子 FLIM 显微镜，该显微镜可为包含青色荧光蛋白（CFP）作为供体和黄色荧光蛋白（YFP）作为受体的 FRET 对提供最佳的 FLIM-FRET 成像。评估和讨论了升级系统的能力，并使用具有代表性的 CFP-YFP FRET 对（mCerulean-mCitrine）的 FLIM-FRET 实验证明了系统的成像性能。