Extended Data Fig. 9: LC-MS analysis of in vitro photoblued AF647.
From: Super-photostable organic dye for long-term live-cell single-protein imaging
a. HPLC chromatography comparison between before (dash) and after photoblueing of AF647 (solid). Mobile phase gradient: H2O with 0.1% TFA (solvent A) and MeOH with 0.1% TFA (solvent B); t = 0-5 min, 10%B hold, t = 5–55 min, 10%B to 50%B; flow rate: 1.2 ml/min; column: C18 column (Phenomenex, 5 μm, 100 Å, 4.6 × 250 mm); column oven: 30 °C. b. positive (left) and negative mode (right) mass spectra of Cy3. Chemical formula of Cy3: C34H44N2O14S4 (calculated m/z: 832.2). c. positive (left) and negative mode (right) mass spectra of peak 1. Chemical formula of peak 1: C35H44N2O15S4 (calculated m/z: 860.2). d. positive (left) and negative mode (right) mass spectra of peak 2. Chemical formula of peak 2: C40H52N2O17S4 (calculated m/z: 960.2). Mass analysis condition: ion source – API-ES; drying gas flow – 11.3 l/min; nebulizer pressure – 35 psi; drying gas temperature – 350 °C; capillary voltage – 3000 V.
