Computational Techniques Toward 1 nm Super-resolution Microscopy

This thesis explores advanced computational techniques in super-resolution microscopy (SRM), with the primary goal of pushing the limits of achievable resolution towards the 1 nm scale. It includes developments in particle fusion algorithms, data analysis of complex biological structures, and exploration of the impact of molecular dipole...

Three-dimensional localization microscopy with increased axial precision through TIRF angle modulation

To better understand the interactions between biological molecules, a high optical resolution in all three dimensions is crucial. The intrinsically lower axial resolution of microscopes however, is a limiting factor in fluorescence imaging, correspondingly in fluorescence based single molecule localization microscopy (SMLM). Here, we present...

Localization microscopy of constrained fluorescent molecules: Pushing towards Ångström-scale resolution through cryogenics

<i>Localization microscopy has circumvented the diffraction limit by sequentially</i> imaging individual light emitting molecules at a time. The position of these individual molecules can be determined and a super-resolution reconstruction is made with improved resolution. Normally freely rotating emitters are used such that the point spread...

Adaptive optics enables multimode 3D super-resolution microscopy via remote focusing

A variety of modern super-resolution microscopy methods provide researchers with previously inconceivable biological sample imaging opportunities at a molecular resolution. All of these techniques excel at imaging samples that are close to the coverslip, however imaging at large depths remains a challenge due to aberrations caused by the...

Completing the canvas: advances and challenges for DNA-PAINT super-resolution imaging

Single-molecule localization microscopy (SMLM) is a potent tool to examine biological systems with unprecedented resolution, enabling the investigation of increasingly smaller structures. At the forefront of these developments is DNA-based point accumulation for imaging in nanoscale topography (DNA-PAINT), which exploits the stochastic and...

Particle fusion in localization microscopy

Single molecule localization microscopy (SMLM) shows promise for quantitative structural analysis of subcellular complexes and organelles with a resolution well below the diffraction limit. This superresolution microscopy technique relies on the blinking events of fluorescent molecules that labeled the structure of interest and are...

Super-resolution imaging visualizes the eightfold symmetry of gp210 proteins around the nuclear pore complex and resolves the central channel with nanometer resolution

One of the most complex molecular machines of cells is the nuclear pore complex (NPC), which controls all trafficking of molecules in and out of the nucleus. Because of their importance for cellular processes such as gene expression and cytoskeleton organization, the structure of NPCs has been studied extensively during the last few decades,...