Multicolour localization microscopy by point-spread-function engineering

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...

Accuracy of the gaussian point spread function model in 2D localization microscopy.

The gaussian function is simple and easy to implement as Point Spread Function (PSF) model for fitting the position of fluorescent emitters in localization microscopy. Despite its attractiveness the appropriateness of the gaussian is questionable as it is not based on the laws of optics. Here we study the effect of emission dipole orientation in conjunction with optical aberrations on the local...