Intravital Fluorescence Excitation in Whole-Animal Optical Imaging.
Intravital Fluorescence Excitation in Whole-Animal Optical Imaging.
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DOI：
10.1371/journal.pone.0149932
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Maitland KC
中科院分区：
文献类型：
--
作者：
Nooshabadi F;Yang HJ;Bixler JN;Kong Y;Cirillo JD;Maitland KC
利用重组或荧光标记的病原体或细胞进行的全动物荧光成像能够实时分析活体动物的疾病进展和治疗反应。组织吸收限制了荧光激发光的穿透，特别是在可见光波长范围内，导致对深层目标的灵敏度降低。在这里，我们演示了使用光纤束将光传递到小鼠肺中以激发荧光细菌，从而避免在整个动物成像中组织吸收激发光。我们提出了使用该技术来提高小鼠肺中表达tdTomato的牛分枝杆菌BCG（卡介苗）细菌的重组报告菌株的检测。将显微内窥镜集成到整个动物荧光成像仪中，以使小鼠肺中的活体激发与整个动物检测成为可能。使用这种技术，检测阈值被测量为103菌落形成单位（CFU）在肺部感染。相比之下，使用标准落射照明的整个动物荧光成像的检测阈值大于106 CFU。
Whole-animal fluorescence imaging with recombinant or fluorescently-tagged pathogens or cells enables real-time analysis of disease progression and treatment response in live animals. Tissue absorption limits penetration of fluorescence excitation light, particularly in the visible wavelength range, resulting in reduced sensitivity to deep targets. Here, we demonstrate the use of an optical fiber bundle to deliver light into the mouse lung to excite fluorescent bacteria, circumventing tissue absorption of excitation light in whole-animal imaging. We present the use of this technology to improve detection of recombinant reporter strains of tdTomato-expressing Mycobacterium bovis BCG (Bacillus Calmette Guerin) bacteria in the mouse lung. A microendoscope was integrated into a whole-animal fluorescence imager to enable intravital excitation in the mouse lung with whole-animal detection. Using this technique, the threshold of detection was measured as 103 colony forming units (CFU) during pulmonary infection. In comparison, the threshold of detection for whole-animal fluorescence imaging using standard epi-illumination was greater than 106 CFU.