Multicolor localization microscopy and point-spread-function engineering by deep learning

Multicolour localization microscopy by point-spread-function engineering.

Super-resolution microscopy has revolutionized cellular imaging in recent years1-4. Methods relying on sequential localization of single point emitters enable spatial tracking at ~10-40 nm resolution. Moreover, tracking and imaging in three dimensions is made possible by various techniques, including point-spread-function (PSF) engineering5-9 -namely, encoding the axial (z) position of a point ...

Point spread function engineering in confocal scanning microscopy

Confocal scanning microscopes are imaging systems that are mainly featured by their unique depth-discrimination capacity when imaging three-dimensional objects. Along the past few years, our research group has done several attempts to improve their axial resolution by means of the so-called point-spread-function (PSF) engineering method. That is, by designing diffractive elements that properly ...

Algorithm for 3d Localization Microscopy Using Realistic Modeling of the Point Spread Function by Phase Retrieval

Programmable vector point-spread function engineering.

We use two nematic liquid crystal spatial light modulators (SLM's) to control the vector point spread function (VPSF) of a 1.3 numerical aperture (NA) microscope objective. This is achieved by controlling the polarization and relative phase of the electric field in the objective's pupil. We measure the resulting VPSF's for several different pupil field polarization states. By using single fluor...

Automated High-speed 3D Multicolor Localization Microscopy

Single-molecule switching nanoscopy (SMSN) provides a powerful tool to investigate structures and dynamics inside the cell at the molecular scale. It is, however, still far from being a routine technology in biology labs. One of the reasons is that 3D multicolor SMSN live-cell imaging still faces significant technical challenges, such as insufficient temporal resolution, high background from ou...