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

单分子定位显微镜（SMLM）及时分离荧光团发射，沿所有三个轴增加十倍的空间分辨率。然而，较长的采集时间使活细胞成像变得困难。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产生人工结构的风险——在三维成像时这种风险进一步增加。Haar小波核软件（HAWK）旨在去除高密度成像过程中的伪影，我们正在使用该技术开发用于SMLM图像的HAWK评估方法（HAWKMAN）。使用各种点扩散函数不对称，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来研究癌细胞在短时间内3D环境中的运动力学。

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.