Middlebury College 1 article published in JoVE Biochemistry Conventional BODIPY Conjugates for Live-Cell Super-Resolution Microscopy and Single-Molecule Tracking Santosh Adhikari1, Chiranjib Banerjee*1, Joe Moscatelli*1,2, Elias M. Puchner1 1School of Physics and Astronomy, University of Minnesota, Twin Cities, Physics and Nanotechnology (PAN), 2Middlebury College Conventional BODIPY conjugates can be used for live-cell single-molecule localization microscopy (SMLM) through exploitation of their transiently forming, red-shifted ground state dimers. We present an optimized SMLM protocol to track and resolve subcellular neutral lipids and fatty acids in living mammalian and yeast cells at the nanoscopic length scale.