Extremely Bright, Near-IR Emitting Spontaneously Blinking Fluorophores Enable Ratiometric Multicolor Nanoscopy in Live Cells
Jonathan Tyson, Kevin Hu, Shuai Zheng, P. Kidd, Neville Dadina, Ling Chu, Derek Toomre, Joerg Bewersdorf, Alanna Schepartz
Abstract
New bright, photostable, emission-orthogonal fluorophores that blink without toxic additives are needed to enable multicolor, live-cell, single-molecule localization microscopy (SMLM). Here we report the design, synthesis, and biological evaluation of Yale676sb, a photostable, near-IR-emitting fluorophore that achieves these goals in the context of an exceptional quantum yield (0.59). When used alongside HMSiR, Yale676sb enables simultaneous, live-cell, two-color SMLM of two intracellular organelles (ER + mitochondria) with only a single laser and no chemical additives.
Topics & Concepts
FluorophoreContext (archaeology)MicroscopyMaterials scienceOrganelleFluorescenceNanotechnologyLaserQuantum yieldBiophysicsChemistryOptoelectronicsOpticsPhysicsBiologyBiochemistryPaleontologyAdvanced Fluorescence Microscopy TechniquesAdvanced Electron Microscopy Techniques and ApplicationsPhotosynthetic Processes and Mechanisms