Parallel excitation-emission multiplexed fluorescence lifetime confocal microscopy for live cell imaging.
Parallel excitation-emission multiplexed fluorescence lifetime confocal microscopy for live cell imaging.
复制标题
DOI：
10.1364/oe.22.010221
复制
发表时间：
2014-05
期刊：
影响因子：
3.8
通讯作者：
Ming Zhao;Yu Li;Leilei Peng
中科院分区：
文献类型：
--
作者：
Ming Zhao;Yu Li;Leilei Peng
我们提出了一种新颖的激发发射多重荧光寿命显微镜 (FLIM) 方法，该方法在多重能力方面超越了当前的 FLIM 技术。该方法采用傅立叶复用以 44,000 像素/秒的速度同时获取多个激发波长和发射颜色组合的共焦荧光寿命图像。该系统采用低成本 CW 激光源和具有多功能光谱配置的标准 PMT 构建，可作为商用共焦显微镜的附加组件。傅里叶寿命共焦方法可实现快速多重 FLIM 成像，从而可以监测活细胞中的多个生物过程。低成本和与商业系统的兼容性也可以使多重 FLIM 更容易被生物研究界所使用。
We present a novel excitation-emission multiplexed fluorescence lifetime microscopy (FLIM) method that surpasses current FLIM techniques in multiplexing capability. The method employs Fourier multiplexing to simultaneously acquire confocal fluorescence lifetime images of multiple excitation wavelength and emission color combinations at 44,000 pixels/sec. The system is built with low-cost CW laser sources and standard PMTs with versatile spectral configuration, which can be implemented as an add-on to commercial confocal microscopes. The Fourier lifetime confocal method allows fast multiplexed FLIM imaging, which makes it possible to monitor multiple biological processes in live cells. The low cost and compatibility with commercial systems could also make multiplexed FLIM more accessible to biological research community.