Intravital imaging of the immune responses during liver-stage malaria infection: An improved approach for fixing the liver.
Intravital imaging of the immune responses during liver-stage malaria infection: An improved approach for fixing the liver.
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DOI：
10.1016/j.parint.2016.02.011
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发表时间：
2016-10
影响因子：
1.9
通讯作者：
M. Akbari;K. Kimura;James T Houts;K. Yui
中科院分区：
文献类型：
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作者：
M. Akbari;K. Kimura;James T Houts;K. Yui
寄主-寄生物关系是现代寄生虫学的主要主题之一。最近的科学革命，包括各种荧光蛋白/探针和双光子显微镜的发展，使得直接可视化和研究宿主与病原体之间相互作用的机制成为可能。在这里，我们描述了我们的方法，准备和建立我们的实验方法，使用活体成像检查伯氏疟原虫ANKA和抗原特异性CD 8 +T细胞之间的相互作用，在肝脏感染阶段的四个维度。由于肝脏位于隔膜附近，因此在成像过程中呼吸运动的中和是至关重要的。此外，由于肝脏的解剖结构和组织结构，血液循环和温度可能会受到手术暴露的影响。为了控制呼吸，我们建议在手术过程中吸入1%的异氟醚进行麻醉。此外，我们的方案在肝脏周围引入了纱布垫，以避免在使用倒置显微镜进行活体成像期间对肝脏的外部压力，这使得可以在血液循环减少最小以及位移和组织损伤最小的情况下对肝脏组织进行长时间成像。该方法的关键是减少活体成像过程中呼吸运动和对肝脏组织的外部压力。
The host–parasite relationship is one of the main themes of modern parasitology. Recent revolutions in science, including the development of various fluorescent proteins/probes and two-photon microscopy, have made it possible to directly visualize and study the mechanisms underlying the interaction between the host and pathogen. Here, we describe our method of preparing and setting-up the liver for our experimental approach of using intravital imaging to examine the interaction between Plasmodium berghei ANKA and antigen-specific CD8+T cells during the liver-stage of the infection in four dimensions. Since the liver is positioned near the diaphragm, neutralization of respiratory movements is critical during the imaging process. In addition, blood circulation and temperature can be affected by the surgical exposure due to the anatomy and tissue structure of the liver. To control respiration, we recommend anesthesia with isoflurane inhalation at 1% during the surgery. In addition, our protocol introduces a cushion of gauze around the liver to avoid external pressure on the liver during intravital imaging using an inverted microscope, which makes it possible to image the liver tissue for long periods with minimal reduction in the blood circulation and with minimal displacement and tissue damage. The key point of this method is to reduce respiratory movements and external pressure on the liver tissue during intravital imaging.