Clear glass coverslip bottom

The TFP probe was tested in cell culture using the HT29 human cancer cell line. HT29 cells were cultured in McCoy’s 5a medium containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin in a black 96-well plate with a clear glass coverslip bottom (Corning Costar). After 24 h, the cell culture medium was replaced with 200 mL of fresh medium containing 5–10 mg of the TFP probe and monitored in a fluorescence microplate reader (Safire II, Tecan) at 37 °C with data points taken every 10 min and then analyzed using Microsoft Excel.

The AD imaging probe was tested in cell culture using the human H4 neuroglioma cell line and H4 cells stably transfected to overproduce BACE1 (H4-BACE1). H4 cells were cultured in Dulbecco’s modified eagle medium (DMEM) containing 10% fetal bovine serum (FBS) (Invitrogen, Carlsbad, CA, USA) and 1% penicillin/streptomycin in a black 96-well plate with a clear glass coverslip bottom (Corning Costar, Corning, NY, USA). H4-BACE1 was cultured in a DMEM medium containing 10% FBS, 1% L-glut/penicillin/streptomycin (Sigma-Aldrich, St. Louis, MO, USA), and 0.5% G418 (EMD Biosciences, San Diego, CA, USA). After 24 hrs, the cell culture medium was replaced with 200 µL of fresh medium containing 5–10 µg of the AD imaging probe and monitored in a fluorescence microplate reader (Safire II, Tecan, Männedorf, Switzerland) at 37 °C with data points taken every 10 min. We used Microsoft Excel to acquire and analyze the data points.