Liu Xiao, Li Ying, Zhu Hui, Zhao Zilong, Zhou Yuan, Zaske Ana-Maria, Liu Li, Li Min, Lu Hujie, Liu Wei, Dong Jing-Fei, Zhang Jianning, Zhang Yanjun
Key Laboratory of Post-trauma Neuro-repair and Regeneration in Central Nervous System, Ministry of Education, Tianjin Key Laboratory of Injuries, Variations and Regeneration of Nervous System, Tianjin Medical University General Hospital, Tianjin Neurological Institute , Tianjin , China .
Platelets. 2015;26(5):480-5. doi: 10.3109/09537104.2014.940888. Epub 2014 Aug 7.
Circulating platelets are anucleated and multi-functional cells that participate in hemostasis and arterial thrombosis. Multiple ligands and mechanical forces activate platelets, leading to cytoskeletal rearrangement and dramatic shape-changes. Such dramatic changes in platelets membrane structures are commonly detected by optical and electron microscopy after platelets are fixed. We have recently developed a method to study the membrane morphology of live platelets using Hopping Probe Ion Conductance Microscopy (HPICM). We have successfully used this technology to study the process of platelet microvesiculation upon exposure to selective agonists. Here, we further discussed technical details of using HPICM to study platelet biology and compared results from HPICM to those from conventional atomic force microscopy and scanning electron microscopy. This method offers several advantages over current technologies. First, it monitors morphological changes of platelets in response to agonists in real time. Second, platelets can be repeatedly scanned over time without damages brought by heat and prolong light exposure. Third, there is no direct contact with platelet surface so that there will no or minimal mechanical damages brought by a cantilever of a conventional atomic force microscopy. Finally, it offers the potential to study platelet membrane ion channels, which have been technically challenging up-to-date. Our data show that HPICM has high-resolution in delineating changes of platelet morphology in response to stimulations and could help to unravel the complex role of platelet in thrombus formation.
循环血小板是无核的多功能细胞，参与止血和动脉血栓形成。多种配体和机械力激活血小板，导致细胞骨架重排和显著的形状变化。血小板固定后，通过光学显微镜和电子显微镜通常可以检测到血小板膜结构的这种显著变化。我们最近开发了一种使用跳跃探针离子电导显微镜（HPICM）研究活血小板膜形态的方法。我们已成功使用该技术研究血小板在暴露于选择性激动剂时的微囊化过程。在此，我们进一步讨论了使用HPICM研究血小板生物学的技术细节，并将HPICM的结果与传统原子力显微镜和扫描电子显微镜的结果进行了比较。该方法相对于现有技术具有几个优点。首先，它实时监测血小板对激动剂的形态变化。其次，血小板可以随时间反复扫描，而不会因热和长时间光照带来损伤。第三，与血小板表面没有直接接触，因此传统原子力显微镜的悬臂不会带来或只会带来最小的机械损伤。最后，它提供了研究血小板膜离子通道的潜力，而这在技术上一直具有挑战性。我们的数据表明，HPICM在描绘血小板形态对刺激的变化方面具有高分辨率，并且有助于揭示血小板在血栓形成中的复杂作用。
