Imaging force-dependent cytoskeletal dynamics in tumour cells using single-molecule localisation microscopy

使用单分子定位显微镜对肿瘤细胞中力依赖性细胞骨架动力学进行成像

• 批准号: 2339463
• 金额: --
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2339463
• 关键词: Imaging; force; dependent; cytoskeletal; dynamics

Single-molecule localisation microscopy (SMLM) separates fluorophore emissions in time to create a tenfold increase in spatial resolution along all three axes. However, long acquisition times make live-cell imaging difficult. These can only be shortened by acquiring 'high-density' data, which increases the risk of generating artificial structures in the final image - a risk that is further increased when imaging in three dimensions. The Haar Wavelet Kernel software (HAWK) is designed to remove artefacts during high-density imaging, and we are using the technology to develop a HAWK Method of Assessment (HAWKMAN) for SMLM images. Using a variety of point-spread function asymmetries, we have also been running evaluations of HAWK's propensity for handling different types of 3D data. Future work will see these technologies applied to live-cell, 3D SMLM imaging using selective plane illumination microscopy to study cancer cell motility mechanics in 3D environments over short timescales.

单分子定位显微镜（SMLM）及时分离荧光团的发射，使空间分辨率沿着所有三个轴增加十倍。然而，长的采集时间使得活细胞成像困难。这些只能通过获取“高密度”数据来缩短，这增加了在最终图像中生成人造结构的风险-当三维成像时，这种风险进一步增加。Haar小波核软件（HAWK）旨在消除高密度成像期间的伪影，我们正在使用该技术开发SMLM图像的HAWK评估方法（HAWKMAN）。使用各种点扩散函数不对称性，我们还对HAWK处理不同类型3D数据的倾向进行了评估。未来的工作将看到这些技术应用于活细胞，3D SMLM成像，使用选择性平面照明显微镜在短时间内研究3D环境中的癌细胞运动机制。