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首页> 外文会议>Conference on photons plus ultrasound: imaging and sensing >Validating tyrosinase homologue melA as a photoacoustic reporter gene for imaging Escherichia coli
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Validating tyrosinase homologue melA as a photoacoustic reporter gene for imaging Escherichia coli

机译：验证酪氨酸酶同系物melA作为用于大肠杆菌成像的光声报告基因

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Antibiotic drug resistance is a major worldwide issue. Development of new therapies against pathogenic bacteria requires appropriate research tools for replicating and characterizing infections. Previously fluorescence and bioluminescence modalities have been used to image infectious burden in animal models but scattering significantly limits imaging depth and resolution. We hypothesize that photoacoustic imaging, which has improved depth-to-resolution ratio, could be useful for visualizing MelA-expressing bacteria since MelA is a bacterial tyrosinase homologue involved in melanin production. Using an inducible expression system, E. coli expressing MelA were visibly black in liquid culture. Phosphate buffered saline (PBS), MelA-expressing bacteria (at different dilutions in PBS), and chicken embryo blood were injected in plastic tubes which were imaged using a VisualSonics Vevo LAZR system. Photoacoustic imaging at 6 different wavelengths (680, 700, 750, 800, 850 and 900nm) enabled spectral de-mixing to distinguish melanin signals from blood. The signal to noise ratio of 9x diluted MelA bacteria was 55, suggesting that ～20 bacteria cells could be detected with our system. When MelA bacteria were injected as a 100 μL bolus into a chicken embryo, photoacoustic signals from deoxy- and oxy- hemoglobin as well as MelA-expressing bacteria could be separated and overlaid on an ultrasound image, allowing visualization of the bacterial location. Photoacoustic imaging may be a useful tool for visualizing bacterial infections and further work incorporating photoacoustic reporters into infectious bacterial strains is warranted.
机译：抗生素耐药性是世界范围内的主要问题。针对病原细菌的新疗法的发展需要适当的研究工具来复制和表征感染。以前，荧光和生物发光方法已用于对动物模型中的传染负荷进行成像，但散射显着限制了成像深度和分辨率。我们假设光声成像具有改进的深度分辨率，可用于可视化表达MelA的细菌，因为MelA是涉及黑色素生产的细菌酪氨酸酶同源物。使用诱导型表达系统，表达MelA的大肠杆菌在液体培养中明显呈黑色。将磷酸盐缓冲盐水（PBS），表达MelA的细菌（在PBS中不同稀释度）和鸡胚血液注入塑料管中，这些管使用VisualSonics Vevo LAZR系统成像。在6个不同的波长（680、700、750、800、850和900nm）上进行光声成像，可以进行光谱混合以区分黑色素信号与血液。 9倍稀释的MelA细菌的信噪比为55，这表明使用我们的系统可以检测到约20个细菌细胞。当将MelA细菌以100μL推注的形式推注到鸡胚中时，来自脱氧和氧合血红蛋白以及表达MelA的细菌的光声信号可以被分离并叠加在超声图像上，从而可视化细菌的位置。光声成像可能是可视化细菌感染的有用工具，因此有必要进一步开展将光声报告基因纳入感染性细菌菌株的工作。

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