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90 protocols using nanowizard 2

1

Atomic Force Microscopy Analysis of Cell Rigidity

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D1 cells (1×10-5) were seeded on 6 cm dishes, incubated for 24 hours, and treated with RhoA stimulator or inhibitor for 30 minutes. Cells were detected using atomic force microscopy (Nano-wizard II; JPK Instruments, Berlin, Germany), and cell rigidity was detected using a cantilever (ArrowTL1; Oxford Instruments, Scotts Valley, CA, USA) with a 5 µm polystyrene sphere. The indentation area targeted cell nuclei. JPK data-processing software was used to analyze cell rigidity according to the Herz model.
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2

Nanoniosome Characterization by AFM and Cryo-TEM

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The shape and the surface morphology of the nanoniosomes were determined using images obtained from an atomic force microscope (AFM) (Nanowizard II; JPK instruments; Germany). The nanoniosome formulations were diluted to 1:1000 with deionized water and sonicated for about 20 mins in an ultrasonic agitator (E–Chrom Tech Co, Taiwan). The preparations were then deposited onto a mica sheet and observed under the AFM. In addition, the bilayer structure of the nanocarriers and its spherical shape were studied by Cryo-TEM (FEI Tecnai 20, type Sphera, OR, USA) at 200 kV.
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3

Nanoparticle Morphology Characterization by AFM

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The morphological characteristics of the nanoparticles were determined at room temperature using an atomic force microscope (Nanowizard II; JPK Instruments; Germany AFM).
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4

Electrode Fabrication and Characterization

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Square-shaped electrodes (500 µm × 500 µm each) were fabricated on glass slides (2 cm × 2 cm × 170 µm) as detailed in the Supplementary Information (Fig. S1a). Prior to use, all electrodes were cleaned by washing with acetone, ethanol, ultrapure water, 10% NaOH aqueous solution at 80 °C (for 15 min), and again ultrapure water. They were studied with AFM (using a NanoWizard II from JPK Instruments) as well as by cyclic voltammetry and EIS carried out using a PGSTAT128N potentiostat (from Metrohm Autolab).
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5

Atomic Force Microscopy of Polyplexes

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Atomic force microscopy (JPK NanoWizard II, Germany) was used to investigate the surface properties, morphology, and sizes of polyplexes. For this, polymers and pDNA were diluted in Tris-HCl buffer (Tris-HCl 10 mM, NaCl 10 mM, MgCl2 2 mM). Then, 10 μl of each solution was placed on a slide, and after fixing, microscopic images of nanoparticles were examined.
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6

Characterizing Nanoparticle Properties by Multimodal Analysis

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The hydrodynamic diameter and surface charge of NPs were measured with a Zetasizer Nano ZS (Malvern Incorporated, Malvern, UK). NPs were dispersed in ultra-pure water, and 20 measurements were conducted per sample. The morphology and distribution of NPs was visualized by TEM (H-7600, Hitachi, Tokyo, Japan) and AFM (NanoWizard II, JPK Instruments, Berlin, Germany). Samples were coated onto carbon film on 200 mesh copper grids (Electron Microscopy Science, Hatfield, PA, USA), dried overnight at room temperature, and visualized by TEM (ARM200F, JEOL, Tokyo, Japan). The levels of AuNP, CPEI, and HP in sCGNPs and mCGNPs were determined by various tools based on the physical and chemical properties of each component. AuNPs were characterized using a UV-vis spectrometer (TECAN PRO 200, Männedorf, Switzerland) in the wavelength range from 450 to 700 nm. CPEI and HP were detected on a fluorescence plate reader (TECAN PRO 200) using a ChemiDoc imaging system (BR170-8265, Bio-Rad Laboratories, Hercules, CA, USA), after which the fluorescence intensity was calculated.
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7

Characterizing CPE45 Nanocrystal Morphology

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All AFM images of CPE45 nanocrystals and single crystals were recorded in tapping mode on a JPK Nano wizard II. For the analysis of nanocrystal thickness, AFM images of nanocrystal films were plane-flattened. A background surface was subtracted, derived from areas showing the bare substrate. A height histogram as shown in Figure 2D was calculated.
The lateral size of the nanocrystals was extracted by measuring cross sections of closely packing nanocrystals. Phase images as shown in Figure 2 were used. The approximate radius of curvature of the AFM-tip represents the experimental error Δx = 10 nm. A bin size of 5 nm for the cross section was used to determine the distribution of particle diameters. For the AFM images of the nanocrystals, AFM-tips with a nominal radius of curvature of 8 or 10 nm and a resonance frequency around 150 kHz or 320 kHz were used. Both had force constants of the cantilever of about 40 N/m. For the images of the functionalized/non-functionalized CPE45 crystals, sharp AFM tips with resonance frequencies of 40–80 kHz and force constants of 0.12 N/m or 0.24 N/m were used.
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8

Perovskite Thin Film Topography Exploration

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Surface topographies of perovskite thin films were inspected using Nano Wizard II (JPK Instruments). The scanning area was 10 μm × 10 μm.
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9

Comprehensive Physicochemical Characterization of Nanomaterials

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The ultrasonic system and injection pump used in this study were a VCX 505 Sonics and a KDS101 syringe pump, respectively (KD Scientific, Holliston, MA, USA). Morphological characteristics were studied with a conventional field emission scanning electron microscope (FE-SEM) (JEOL JSM-7500F, 15 kV) and a biological atomic force microscope (Bio-AFM) (NanoWizard II; JPK) in intermittent air mode. Ultraviolet (UV)-visible spectra were analyzed with an Optical 3220. X-ray diffraction (XRD) (Scintag-SDS 2000) was performed at 40 kV voltage and 30 mA current with continuous-scanning 2θ mode. Fourier transform infrared (FT-IR) spectra of the samples were recorded at room temperature with a TENSOR 27 instrument (Bruker) in the 650–4,000 cm−1 region. CV measurements were recorded with VersaSTAT 3 (VersaStudio) in a three-electrode configuration. Zetasizer Range (Nano-Z; Malvern Instruments, Malvern, UK) was used for particle size measurement. The stained samples were analyzed using laser scanning fluorescence microscopy (Nikon Eclipse TE 2000-U) and fluorescence-activated cell sorting (FACS) (BD FACSCalibur™; BD Biosciences, San Jose, CA, USA).
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10

Evaluating PDMS Mechanical Properties

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To evaluate the Young's modulus of the PDMS, stress–strain experiments were carried out at room temperature with a tensile machine (Instron model 5542). PDMS rectangular arrays of 26 mm in perimeter were tested at an initial strain rate of 2 mm min–1. The slope at the start of the deformation was used to calculate the Young's modulus E. We found that by curing PDMS at 65 °C for 6 h, the E was 3±0.3 MPa. After measuring the micropillar dimensions by SEM, we calculated micropillar spring constants according to:

where I is the moment of inertia and L the length of the micropillar61 . Alternatively, we used an AFM (Nanowizard II, JPK Instruments) to directly evaluate the spring constant of shorter micropillars with lengths of 8.2 μm and of longer micropillars with lengths of 13.7 μm (Supplementary Fig. 2). To do this, the top of a micropillar was brought into contact with an AFM cantilever. The AFM cantilever was used to apply a force to a micropillar and the micropillar deflection was measured. From this deflection, we determined the average spring constant of the shorter micropillars.
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