Data processing software

The microemulsions morphology was studied by atomic force microscopy (AFM) (JPK, Nano Wizard 2, and Berlin Germany). For AFM analysis, samples were diluted by deionized water and then the clean mica surface were smeared by a drop of the diluted sample, followed by being lyophilized to prepare a fixed sample. All AFM images were processed and analyzed using the JPK Data Processing software.

To estimate the size of protruding bulges on the cell membranes, 10 × 10 µm fields on the surface of different ADSCs were randomly selected and scanned by AFM. Images were flattened by 9^th grade polynomial fit subtraction on each scan line independently, using JPK Data Processing software. In this way, cell surface slopes not related to protrusions can be filtered in the analysis. Using Wasabi! software, cross-area above a fixed height threshold was measured on each image, in order to produce a value representing the amount of protrusions in that field.

Samples were prepared on freshly cleaved mica and
dried at room temperature.
AFM images were acquired using a commercial AFM system (JPK NanoWizard
3 and 4). Measurements were performed in AC mode with SNL-10 probes
(Bruker) at 25 °C, 35–40% RH. AFM images were collected
with 1024 × 1024 pixels/frame. Each AFM tip was characterized
prior to usage. Analyses of AFM images were performed with JPK Data
Processing software. Note that for the height analyses of the AFM
images, the baseline height was leveled against the flat base plane
of the substrate. All AFM images were only subjected to the primary
first order flattening correction to remove sample tilt so that potential
artifacts induced by other image processing steps were avoided as
much as possible.

Mechanical properties of BD PuraMatrix Peptide Hydrogel (Corning, Tewskbury, MA, USA) were investigated using atomic force microscopy (AFM, CellHesion head, JPK Instruments, Berlin, Germany) in force mapping mode. To probe hydrogel samples, commercially-available silicon nitride cantilevers (MLCT-C, Bruker, Billerica, MA, USA) with a nominal spring constant of 0.01 N/m were applied. Force curves, i.e., dependencies between cantilever deflection and relative sample position, were acquired over a grid of 8 × 8 pixels within a scan area of 50 × 50 μm. The maximum load force (F) was 5 nN and load speed of 8 μm/s was maintained. Young’s modulus was determined using Hertz contact mechanics as described previously [77 (link)]. Briefly, the following relation between the load force (F) and resulting indentation (Δz) for the paraboloidal assumption of the probing tip was applied: $F=\frac{4·\sqrt{R}·E}{3·\left(1-v^2\right)}·\Delta z^{\raisebox{1ex}{$3$}\!\left/ \!\raisebox{-1ex}{$2$}\right.}$
JPK Data Processing software was used to apply this equation to the experimental data to obtain the value of Young’s modulus for each force curves. The final Young’s modulus was obtained by averaging all force curves and was expressed as a mean and standard deviation.