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

单分子定位显微镜(SMLM)及时分离荧光团排放，沿所有三个轴的空间分辨率提高十倍。然而，长时间的采集使活细胞成像变得困难。只有通过获取高密度数据才能缩短这些时间，因为高密度数据会增加在最终图像中生成人造结构的风险--在三维成像时，这种风险会进一步增加。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为SMLM图像开发一种HAWK评估方法(Hawkman)。利用各种点扩散函数的不对称性，我们还对霍克处理不同类型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，使用选择性平面照明显微镜进行3D SMLM成像，以在短时间尺度上研究3D环境中癌细胞的运动机制。

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.