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A custom Software for the Evaluation of Single-Cell Force-Spectroscopy Data Acquired by FluidFM BOT

FluidFM technology is a novel atomic force microscopy (AFM) based tool with microfluidic and pressure control extensions. One of its main features is the robotized version called the FluidFM BOT, which can perform up to 30-40 Single-Cell Force-Spectroscopy (SCFS) measurements per day exceeding previous SCFS acquisition methods. The user software of the FluidFM BOT allows the user to measure the inverse optical lever sensitivity (InvOLS) and spring constant (k) of the applied FluidFM micropipette cantilevers used for SCFS recording, which parameters directly scale the obtained voltage values from the two-segmented photodetector. However, the user software miscalculates the InvOLS values and does not allow exporting measurement parameters Fmax, Emax, and Dmax, which are characteristic of individual SCFS Force-Distance (FD) curves. Also, exporting individual FD-curve plots is not possible. Here we show a software specially developed in Matlab for the correct acquisition of InvOLS values, plotting and exporting of FD-curves and their parameters. The software allows users to interfere, but it is partially automated to enable smooth data evaluation procedures. The software will be available free of charge on the link provided at the end of the manuscript.