Date of Award

Spring 2022

Document Type


Degree Name

Doctor of Philosophy (PhD)


Biomedical Engineering (ENAS)

First Advisor

Bewersdorf, Joerg


plays a critical role in cell biology due to its ability tovisualize molecules of interest inside living cells with high specificity and contrast. However, diffraction of light limits spatial resolution to ~250 nm. Super-resolution microscopy bypasses this physical limit to achieve resolutions of ~10 nm, which is nearly the size of many biological macromolecules. Notably, 4Pi single-molecule switching nanoscopy achieves this resolution in all three spatial dimensions, finally making it possible to resolve specific proteins within convoluted structures like the Golgi. However, the current implementation is only compatible with dead or fixed cell samples, restricting experimental design. To explore the complexity of life in 3-D at near molecular resolution, I have developed a 4Pi single-molecule switching nanoscope for multi-color live-cell imaging and optimized 2-color live-cell labeling with spontaneously-blinking fluorophores.