Poly (ADP-ribose) polymerase 1 is a key player in DNA repair and transcription regulation. The role of PARP1 in DNA damage recognition and protein signaling in response make it integral to genome stability. Beyond this, it regulates gene expression through chromatin remodeling and transcription factor binding. Many of these activities are dependent upon PARP1 autoPARylation, induced by DNA binding. PARP1 has long been implicated in G-quadruplex DNA recognition, and recent advancements in detection of these DNA structures in living tissue have shown that we barely understand PARP1 function and activity. In the first part of this thesis, I will describe single-molecule TIRF microscopy experiments that identified PARP1: DNA complexes vary by DNA structure. Next, I will describe my work to modify a new technique to characterize PARP1 DNA binding and autoPARylation on over 10,000 DNA G-quadruplexes. By combining next generation sequencing and single-molecule TIRF microscopy, I demonstrate the power of this massively parallel single-cluster TIRF method. Finally, I will make the case for the need to develop this methodology to the level of routine lab experiment as it can have profound impacts on advancing our understanding of the influences the physical landscape of the genome has on cellular processes.