| Summary: | 碩士 === 國立陽明大學 === 生醫光電研究所 === 106 === Single-particle tracking (SPT) is of growing importance in the biophysical community. SPT allows researchers to follow the fate of individual virus particles, to probe dynamic interactions between viruses and cellular machinery. In the past, our lab’s publication also use epi fluorescence microscopy to observe two-dimensional tracking images. Reveals the relationship between dengue virus and autophagosome in the early stage of infection. However, intracellular is a three-dimensional space, and it is necessary to develop three-dimensional real-time single particle tracking.
In order to upgrade the system to the 3D level, we have used an electrically tunable lens that can quickly change the imaging plane. The electrically tunable lens is mounted under the objective lens to synchronize the excitation light and the radiation energy by ETL regulation. And write the LabView program synchronization CMOS, shutter and electronically controlled variable lens connection. After measuring the z-axis of the dried fluorescent beads, it is known that the z-axis distance varies by about 0.13 μm per 1 mA of the ETL. During the one hour of tracking the dried fluorescent beads, the z-axis position and the fluorescence intensity of the fluorescent beads remained unchanged, indicating the stability of the system. In addition, the system effectively tracks the movement of the fluorescent beads as the dried fluorescent beads move over the z-axis over time. And we used dengue virus to infect Huh7.5 cells expressing GFP-LC3, and tracked the movement of the organelles and virus particles in living cells. In summary, the system achieves a z-axis resolution of at least 10nm and a time resolution of less than 1 second, and enables image and tracking of live cells and virus particles.
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