Nanowizard 2 afm

All solvents and starting materials for synthesis were commercially available and were used as received. Fourier transform infrared spectra (FTIR) were collected using a Shimadzu IRAffinity-1 from 4000–400 cm⁻¹. PXRD patterns were collected at room temperature using an Empyrean PANalytical diffractometer (CuKα_1,2 radiation, λ₁ = 1.540598 Å and λ₂ = 1.544426 Å) equipped with a PIXcel 1D detector and a flat-plate sample holder in a Bragg–Brentano para-focusing optics configuration (45 kV, 40 mA). Intensity data were obtained by the step counting method (step: 0.02°) in continuous mode in the approximate range 3.0° ≤ 2θ ≤ 50°. Scanning electron microscope (SEM) images were acquired using a Philips CM-200 (200 kV). Fluorescence spectra for the characterization of the MOF and for DNT sensing were collected using a Hitachi F-7000 Fluorescence Spectrophotometer (Hitachi High Technologies, Krefeld, Germany). Fluorescence spectra of films containing [Zn₂(bpdc)₂(bpee)] were obtained by using a sample holder for solid samples. AFM images were acquired by means of a JPK NanoWizard II AFM functioning in contact mode, connected to a Nikon Eclipse Ti inverted optical microscope. MikroMasch HQ:XSC11/Al BS cantilevers of 0.2 N/m spring constant and 15 kHz resonant frequency were used in air conditions.

AFM was performed in a JPK NanoWizard II® AFM coupled to a Nikon Eclipse Ti-U inverted optical microscope. Dynamic mode immersed in liquid with an Olympus commercial silicon nitride cantilever tip (0.76 N/m, 71 kHz), with typical 15 nm radius at the end, was used. In vivo analyses were performed at 37 °C constant temperature maintained by a JPK BioCell thermocontroller. AFM characterizations on fixed cells were done after adding equivalent amounts of paraformaldehyde 4% to the medium for 10 min and changing to glutaraldehyde 2.5% for 2–3 h. For the study of the effect of drugs on the generation of cell protrusions, 0.5 mM progesterone (water soluble, Sigma-Aldrich), 10 ng/ml PDGF-BB or 10 ng/ml PDGF-AA (both from ImmnoTools GmbH) were added to the MesenPRO medium that had been changed 1 h before so that pH and temperature were stabilized at the moment of the treatment. Experimental conditions also included incubation in defined medium (α-MEM, Sigma-Aldrich) that had been pre-warmed and pre-equilibrated for pH. The cells were fixed, as above, at different times after the addition of drugs or medium change.