KELLY neuroblastoma cells were cultured in Corning RPMI 1640 (Millipore Sigma, Burlington, MA) supplemented with 100 U/mL penicillin and 100 μg/mL streptomycin (Fisher Scientific), 5 mL reconstituted oxaloacetate-pyruvate-insulin (Millipore Sigma), 10% Foundation B fetal bovine serum (Gemini Bioproducts, West Sacramento, CA), and 2 mM L-glutamine (Fisher Scientific). KELLY cells were incubated 24 hours at 37°C in 5% CO2 in humidity to allow surface adhesion before ICG administration. IR-125 laser grade indocyanine green (ICG) was purchased from Fisher Scientific (Waltham, MA). Bovine serum albumin (BSA) was purchased from Millipore Sigma (St. Louis, MO).
For proliferation and viability tests, cells were seeded in a 96-well plate at a density of 12,500 cells/cm2, and were incubated for 24 hours with aqueous ICG, photobleached ICG, or no ICG (control). 20 μL of 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS; Promega Corporation, Madison, WI) was added to each well and incubated for 3 hours for each data point. MTS in complete RPMI absent of cells served as a blank for each timepoint. Proliferation data was normalized to the timepoint before treatment. Viability was determined by comparing MTS absorbance of treated cells by normalizing the absorbance of untreated cells tested simultaneously.
For proliferation and viability tests, cells were seeded in a 96-well plate at a density of 12,500 cells/cm2, and were incubated for 24 hours with aqueous ICG, photobleached ICG, or no ICG (control). 20 μL of 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS; Promega Corporation, Madison, WI) was added to each well and incubated for 3 hours for each data point. MTS in complete RPMI absent of cells served as a blank for each timepoint. Proliferation data was normalized to the timepoint before treatment. Viability was determined by comparing MTS absorbance of treated cells by normalizing the absorbance of untreated cells tested simultaneously.