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.
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