The double helix PSF: implementation and localization applications

Valentin Oreiller

Semester Project

Section of Life Sciences and Technologies, EPFL

July 2013

Abstract

Super resolution microscopy technologies like STORM, PALM and fPALM are useful tools to study the structure and the behavior of cells at nanometer scale. The principle underlying these methods is to photo-activate a randomly distributed subset of fluorophores at different time frames and summing up lit locations to compose an image. The localization of the point sources is done using a point spread function (PSF). One way to improve the axial localization is to use a phase mask which generates double-helix PSFs. It means that the PSFs exhibit two lobes which rotate in function of the depth particle. The aim of this project is to implement such a phase mask and to optimize it.

defocusedPSF.avi