Shadowless-illuminated variable-angle TIRF (siva-TIRF) microscopy for the observation of spatial-temporal dynamics in live cells
作者： Weijian Zong , Xiaoshuai Huang , Chi Zhang , Tianyi Yuan , Ling-ling Zhu
DOI: 10.1364/BOE.5.001530
关键词:
摘要: Total-internal-reflection fluorescence (TIRF) microscopy provides high optical-sectioning capability and a good signal-contrast ratio for structures near the surfaces of cells. In recent years, several improvements have been developed, such as variable-angle TIRF (VA-TIRF) spinning (sp-TIRF), which permit quantitative image analysis address non-uniform scattering fringes, respectively. Here, we present dual-color DMD-based shadowless-illuminated (siva-TIRF) system that uniform illumination field. By adjusting incidence angle illuminating laser on back focal plane (BFP) objective, can rapidly illuminate biological samples in layers various thicknesses or hollow-cone epi-fluorescence mode. Compared with other methods accomplishing VA-TIRF/sp-TIRF illumination, our is simple to build cost-effective, it optimal multi-plane images. showing spatiotemporal correlated movement clathrin-coated microtubule filaments from live cells, demonstrate cortical microtubules are important spatial regulators structures. Moreover, be used prove superb axial information three-dimensional plasma membrane within
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