Multiflo fx multi mode dispenser

Cells were seeded in 384-well plates at 1,000 cells/well in 40 µL of growth medium. After 24 hours, cells were treated with varied concentrations of drugs from a small Food and Drug Administration (FDA)-approved cancer drug library consisting of 62 cancer drugs (purchased from Selleck Chemicals or MedChemExpress) with 28 specific targeted molecules for 5 days. Cell numbers were determined by the Operetta^® High Content Screening System (PerkinElmer, Waltham, MA, USA). The images were used to calculate growth rate inhibition (GR) values using the following equation (18 (link)).
For high-throughput drug screening, cells were seeded in 384-well plates using a MultiFlo FX Multimode Dispenser (BioTek Instruments, Winooski, VT, USA) at 1,000 cells/well in 40 µl of growth medium. After 24 hours, cells were treated with the drugs in the drug library (at their respective GR₇₅ values) in 20 µl of the total growth medium and using an EpMotion pipetting robot (Eppendorf, Hamburg, Germany). The cells were incubated for 5 days in a 37°C 5% CO₂ environment. Cells were stained with 4,6-diamidino-2-phenylindole (DAPI) and imaged using an Operetta CLS™ high-content system (PerkinElmer). The nuclei number was analyzed using a Columbus Image Data Storage and Analysis System (PerkinElmer), and plotted using MATLAB 2017a software (MathWorks, Natick, MA, USA).