Doctoral Defence S.T. Hung

Super-resolution microscopy in tissue through illumination engineering and aberration correction

 

Promotor:  Prof.dr.ir. Michel Verhaegen

Co-promotor: Dr.ir. Carlas Smith, Prof.dr. Nynke Dekker

Abstract: Super-resolution microscopy has been demonstrated to surpass the diffraction limit, delivering high-resolution images. State-of-the-art super-resolution research continually strives to improve imaging resolution using existing super-resolution technology. This thesis aims to enhance super-resolution microscopy resolution through engineered illumination and establish a solid theoretical foundation to support resolution improvements.

In Chapter 2, I discuss the enhancement of resolution in single-molecule localization microscopy for thick sample imaging. In thick samples, the issue of high background noise can degrade the localization precision of single-molecule localization. We introduce Single Objective Lens Lightsheet Microscopy (SOLEIL) as a solution to mitigate the high background photon issue, thereby improving the resolution of single-molecule localization microscopy. Furthermore,  in Chapter 3, I propose an imaging solution to address the high background issue and sample-induced aberration in thick sample imaging. In this chapter, I suggest combining SOLEIL microscopy with a sensorless Adaptive Optics (AO) method to address these issues. In Chapter 4, I propose a comprehensive PSF model theory for Image Scanning Microscopy (ISM) that fully considers vectorial effects, aberrations, Stokes shifts, and Fresnel coefficients at media interfaces. This approach allows for the accurate modeling of excitation and emission PSFs, providing precise references for selecting hardware parameters and assessing image quality.