Point spread function engineering for fluorescence microscopy

Abstract

ENGLISH SUMMARY: A tool for biological imaging is developed within this work. It consists of engineering the point spread function (PSF) of a fluorescent molecule or a nanoparticle emitter by modulating the phase of the fluorescence emission. The engineered PSF developed in this current work is called the double helix point spread function (DH-PSF). Information about the three dimensional position of an emitter and its orientation can be extracted using this PSF with a very high precision and accuracy. Other modalities, such as the pyramidal PSF and the bisected PSF, are also simulated and compared theoretically with the efficiency of the DH-PSF.

AFRIKAANSE OPSOMMING: ‘n Afbeeldingstegniek vir biologiese materie word in hierdie projek ontwikkel. Die tegniek behels die verstelling van die puntverspreidingsfunksie (PVF) van ‘n fluoresserende molekuul of nanopartikel deur die fase van die emissie te moduleer. In die huidige projek word daar gefokus op ‘n spesifieke PVF genaamd, dubbel helix puntverspreidingsfunksie (DH-PVF). Deur die korrekte toepassing van die DH-PVF is dit moontlik om hoë presisie en akkurate drie dimensionele posisie asook orientasie inligting van enkel fluoresserende voorwerpe te bepaal. Verder word daar ook ander modaliteite naamlik piramidale PVF en verdeelde PVF gesimuleer en met die DH-PVF vergelyk.