1. 제목: Correlative Microscopy for 3D Structural Analysis of Dynamic Interactions
2. 연사: 전상미 선임 연구원 ( 한국기초과학지원연구원 질환표적기능연구실 )
3. 일시: 2016. 6. 1 (Wed) PM 4:30 ~ 5:45
4. 장소: #219, CHUNG Moon Soul Bldg.
5. 초록
Cryo-electron tomography (cryoET) allows 3D visualization of cellular structures at molecular resolution in a close-to-physiological state. However, due to the poor signal, low contrast, and radiation sensitive nature of unstained frozen-hydrated specimens, acquisition of tomographic projection series is not generally selective in choice of imaging targets. Therefore, the full potential of cryoET for 3D cellular imaging is realized especially for cellular processes that are rare or dynamic. In order to overcome this limitation in cryoET analysis, approaches for correlating fluorescent light microscopy and cryoET are highly desirable, not only to complement the structural information obtained from cryoET with the dynamic functional data from fluorescent labeling, but also to guide sampling in cryoET. Such tools are particularly valuable for investigating the early events of HIV-1 infection in cells, which are infrequent and difficult to catch. Here, we report on a methodology that combines high speed 3D live-cell imaging with cryoET tools. We applied this technology to visualize the process viral entry into HeLa cells, following of the same particles. Through direct 3D visualization, we identified individual, stationary HIV-1 particles (~ 100 nm in size) bound to the cell surface is located near filamentous structures, whereas moving particles were found to be closely associated with the cytoplasmic intracellular trafficking network. We showed, for the first time under near-native conditions, that intact hyperstable mutant HIV-1 cores are released into the cytoplasm of host-cells. Taken together, we anticipate that the methodology established here will not only constitute a useful tool for studying virus-host cell interactions at various stages during infection, but will also open new ways to investigate cell signaling events and many other cellular processes in general.

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