3D microscopy in extreme conditions: in-flow or at high density

Attaining three-dimensional data at high throughput is a grand challenge in microscopy. I will discuss two recent contributions to 3D microscopy utilizing point-spread-function (PSF) engineering: 1.a new method that extends the capabilities of flow-based imaging to 3D co-localization microscopy, and 2. efficient and fast localization of dense molecules in volumetric single-molecule-localization microscopy (SMLM) using deep learning, termed DeepSTORM3D. Together, these methods enable the study of sub-cellular biological processes at unprecedented timescales and throughputs.