96 pin woundmaker tool

Cells were plated at 35,000 cells per well in Image Lock 96 well plates (Essen BioScience) in 100 μL complete DMEM medium. After overnight incubation, a scratch was made in the plate using a 96-pin WoundMaker tool (Essen BioScience). Plates were washed and incubated for 1 h, and then appropriately diluted drugs were added in 100 μL media to achieve final concentrations. Plates were scanned with an Incucyte Zoom (Essen BioSciences) according to manufacturer's instructions at 2-hour intervals. Images were automatically acquired and registered by the IncuCyte™ software system (ESSEN BioScience). Typically, kinetic updates were recorded over a 24 h period.

30,000 HUVEC were seeded on ImageLock 96-well plates (Essen Bioscience). Once a monolayer had formed, this was scratched the following day with a 96-pin WoundMaker tool (Essen Bioscience). EGM was then refreshed to remove dead and scraped cells. Afterwards, the cells were imaged in an Incucyte imaging system for 11 h (one image every one hour, with the “phase” image channel and 10× magnification). The Scratch Wound Cell Migration Module of the Incucyte S3 Live Cell Analysis System (Essen Bioscience) was used to monitor and analyze the cells.