3D-resolved fluorescence and phosphorescence lifetime imaging using temporal focusing wide-field two-photon excitation
3D-resolved fluorescence and phosphorescence lifetime imaging using temporal focusing wide-field two-photon excitation
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DOI：
10.1364/oe.20.026219
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发表时间：
2012-11-19
期刊：
影响因子：
3.8
通讯作者：
So, Peter T. C.
中科院分区：
文献类型：
--
作者：
Choi, Heejin;Tzeranis, Dimitrios S.;So, Peter T. C.
荧光和磷光寿命成像是研究细胞内蛋白质相互作用和诊断组织病理生理学的有力技术。虽然寿命分辨显微镜长期以来一直是生物光子学社区的剧目，但目前的实现在同时提供3D分辨率，高通量和良好的组织穿透方面存在不足。该报告描述了一种新的高效的寿命分辨成像方法，该方法结合了时间聚焦宽场多光子激发和使用纳秒选通成像仪从3D分辨平面同时采集频域中的寿命信息。该方法是可扩展的，允许仅受可用激光峰值功率限制的快速体积成像。所提出的方法的准确性和性能证明在几个重要的成像研究了解周围神经再生过程。最重要的是，这种方法的并行性可以将长寿命过程（例如磷光）的成像速度提高几个数量级。(C)2012美国光学学会
Fluorescence and phosphorescence lifetime imaging are powerful techniques for studying intracellular protein interactions and for diagnosing tissue pathophysiology. While lifetime-resolved microscopy has long been in the repertoire of the biophotonics community, current implementations fall short in terms of simultaneously providing 3D resolution, high throughput, and good tissue penetration. This report describes a new highly efficient lifetime-resolved imaging method that combines temporal focusing wide-field multiphoton excitation and simultaneous acquisition of lifetime information in frequency domain using a nanosecond gated imager from a 3D-resolved plane. This approach is scalable allowing fast volumetric imaging limited only by the available laser peak power. The accuracy and performance of the proposed method is demonstrated in several imaging studies important for understanding peripheral nerve regeneration processes. Most importantly, the parallelism of this approach may enhance the imaging speed of long lifetime processes such as phosphorescence by several orders of magnitude. (C) 2012 Optical Society of America