Abstract
This protocol introduces the SuperSTORM technique, combining stochastic optical reconstruction microscopy (STORM) and molecular modeling. SuperSTORM is optimized for acquiring and processing STORM images of neutrophil integrins but can be used for any cell-surface molecule with known structure and antibody-binding site(s). SuperSTORM identifies molecular cut-offs for eliminating multiple blinks of STORM imaging, determines colocalization, identifies clusters, and reveals molecular orientations and distributions. This protocol extends STORM imaging to cells in microfluidic systems. Improved resolution is achieved by using biomolecule-inherent parameters. For complete information on the generation and use of this protocol, please refer to the paper by Fan et al. (2019).