Kling Chantal, Smith Todd G, Baird Nicolle, Hutson Christina L
Poxvirus and Rabies Branch, Centers for Disease Control and Prevention, Atlanta, GA, USA.
Methods Mol Biol. 2025;2860:175-189. doi: 10.1007/978-1-0716-4160-6_12.
Bioluminescent images of viral replication in live animals (in vivo) reveal disease dynamics and effects of medical countermeasures over time. After selecting an appropriate orthopoxvirus animal model for the study, a recombinant virus with the firefly luciferase gene inserted in the genome is used to infect the animals. On the day of bioluminescent imaging, the substrate, D-luciferin, is prepared; animals are sedated and injected with the substrate and IVIS imager is utilized; various bioluminescent images are acquired; then animals recover and are able to continue in the study. Ex vivo imaging can also be completed after animals are euthanized at experimental endpoint. This approach allows real-time imaging of viral kinetics within an animal, and analysis of images can provide an additional quantitative measure throughout the study. Bioluminescent imaging not only provides scientific benefits but also benefits to animal welfare. For these reasons, bioluminescent imaging should be considered for any in vivo orthopoxvirus study.
活体动物（体内）病毒复制的生物发光图像揭示了疾病动态以及医学应对措施随时间的效果。在为该研究选择合适的正痘病毒动物模型后，使用一种基因组中插入了萤火虫荧光素酶基因的重组病毒来感染动物。在进行生物发光成像的当天，准备底物D - 荧光素；对动物进行镇静并注射底物，然后使用IVIS成像仪；采集各种生物发光图像；接着动物恢复并能够继续参与研究。在实验终点对动物实施安乐死后，也可以完成离体成像。这种方法能够对动物体内的病毒动力学进行实时成像，并且在整个研究过程中，对图像的分析可以提供额外的定量测量。生物发光成像不仅具有科学价值，还对动物福利有益。基于这些原因，对于任何体内正痘病毒研究都应考虑采用生物发光成像技术。
