Singlet oxygen phosphorescence lifetime imaging based on a fluorescence lifetime imaging microscope.
Singlet oxygen phosphorescence lifetime imaging based on a fluorescence lifetime imaging microscope.
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DOI：
10.1021/acs.jpca.5b01504
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发表时间：
2015-03
期刊：
影响因子：
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通讯作者：
Wenming Tian;Liezheng Deng;Shengye Jin;Heping Yang;Rongrong Cui;Qing Zhang;Wenbo Shi;Chunlei Zhang;Xiaolin Yuan;G. Sha
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文献类型：
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作者：
Wenming Tian;Liezheng Deng;Shengye Jin;Heping Yang;Rongrong Cui;Qing Zhang;Wenbo Shi;Chunlei Zhang;Xiaolin Yuan;G. Sha
在改进的荧光寿命成像显微镜（FLIM）上研究了单线态氧磷光（SOP）寿命成像显微镜的可行性。O2(a(1)Δg→X（3)Σg(-)）和O2(a(1)Δg在C60粉末样品中通过光敏化工艺产生红外辐射跃迁的SOP结果。为了捕获非常微弱的SOP信号，在FLIM的物镜和管透镜之间放置了一个二向镜，用于将样品返回的发光分为两束：反射的SOP光束和C60 （C60- pl）光束的透射光致发光。C60-PL光束进入FLIM的扫描仪，沿着FLIM的正常光路，而SOP避开扫描仪，直接进入精心设计的SOP检测通道。然后采用激光扫描方式同时获得共聚焦C60-PL图像和非共聚焦SOP图像。实验结果表明：(1)在激光扫描模式下，共焦SOP成像的障碍是红外不兼容扫描仪，可以通过使用红外兼容扫描仪来解决。同时在SOP检测通道中增加一个针孔，在阶段扫描模式下也有望实现共聚焦SOP成像。(2) SOP成像的一大挑战是其成像时间过长，从荧光图像中只选择几个有趣的点来测量其SOP随时间变化的轨迹可能是一个正确的折衷方案。
The feasibility of singlet oxygen phosphorescence (SOP) lifetime imaging microscope was studied on a modified fluorescence lifetime imaging microscope (FLIM). SOP results from the infrared radiative transition of O2(a(1)Δg → X(3)Σg(-)) and O2(a(1)Δg) was produced in a C60 powder sample via photosensitization process. To capture the very weak SOP signal, a dichroic mirror was placed between the objective and tube lens of the FLIM and used to divide the luminescence returning from the sample into two beams: the reflected SOP beam and the transmitted photoluminescence of C60 (C60-PL) beam. The C60-PL beam entered the scanner of the FLIM and followed the normal optical path of the FLIM, while the SOP steered clear of the scanner and directly entered a finely designed SOP detection channel. Confocal C60-PL images and nonconfocal SOP images were then simultaneously obtained by using laser-scanning mode. Experimental results show that (1) under laser-scanning mode, the obstacle to confocal SOP imaging is the infrared-incompatible scanner, which can be solved by using an infrared-compatible scanner. Confocal SOP imaging is also expected to be realized under stage-scanning mode when the laser beam is parked and meanwhile a pinhole is added into the SOP detection channel. (2) A great challenge to SOP imaging is its extraordinarily long imaging time, and selecting only a few interesting points from fluorescence images to measure their SOP time-dependent traces may be a correct compromise.