用于定量钙离子成像的基因编码荧光共振能量转移（FRET）传感器的共聚焦荧光寿命成像（FLIM）

Confocal FLIM of genetically encoded FRET sensors for quantitative Ca2+ imaging.

作者信息

Sauer Benjamin, Tian Qinghai, Lipp Peter, Kaestner Lars

机构信息

Institute for Molecular Cell Biology and Research Center for Molecular Imaging and Screening, School of Medicine, Saarland University, 66421 Homburg/Saar, Germany.

出版信息

Cold Spring Harb Protoc. 2014 Dec 1;2014(12):1328-32. doi: 10.1101/pdb.prot077040.

DOI:10.1101/pdb.prot077040

PMID:25447281

Abstract

Fluorescence lifetime imaging (FLIM) is a powerful imaging mode that can be combined with confocal imaging. Changes in the fluorescence decay time of a donor in an intramolecular Förster resonance energy transfer (FRET)-based biosensor provide intrinsic quantitative data. Here, we describe a protocol using both the Ca(2+) sensor TN-XL, which uses troponin C, as the Ca(2+)-sensing unit, and the FLIM technology based on time-correlated single-photon counting.

摘要

荧光寿命成像（FLIM）是一种强大的成像模式，可与共聚焦成像相结合。基于分子内Förster共振能量转移（FRET）的生物传感器中供体荧光衰减时间的变化可提供内在定量数据。在此，我们描述了一种使用Ca(2+)传感器TN-XL（其使用肌钙蛋白C作为Ca(2+)传感单元）和基于时间相关单光子计数的FLIM技术的方案。