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26 protocols using nanoscope 5

1

Evaluating Cellulose Nanofiber Morphology

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The cellulose nanofibers and the modified versions were examined using AFM to understand the morphology of the materials. The samples were prepared by dispersing the freeze-dried nanofibers in water and drop-casting from dilute suspension on mica. An atomic force microscope (AFM) Nanoscope V from Veeco Instruments Inc., Plainview, NY, USA, at a resonance frequency of 310 kHz was used and the nanofiber sizes were evaluated from the height images using Nanoscope V software.
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2

Nanoscale DNA Structure Analysis Using AFM

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AFM measurements were performed at room temperature using Dimension 3100 D31005-1
with Nanoscope V (Veeco). AFM images were recorded on freshly cleaved mica
surfaces (TED PELLA, Inc.). A 10-μl aliquot of the solution containing
the DNA nanostructures was deposited in the presence of 10 mM
Mg(Ac)2. The surfaces were rinsed with 10 mM
Mg(Ac)2 solution and dried under a stream of air. Images were
recorded with AFM tips (Model NSC11, Umasch, and Model RTESP, Part MPP-11100-10,
BRUKER) and using tapping mode at their resonance frequency. The images were
analysed using NANO Scope analysing software (Vecco). The nanostructures chosen
for evaluations were auto-selected and analysed using NANO Scope analysing
software (Vecco). More specifically, all particles with a minimum size of
0.5 nm and a maximum size of 3 nm were auto-selected from the AFM
images.
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3

Microgel Size Characterization by AFM

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The AFM measurements (Bruker, Billerica, USA) were done in the tapping mode using a Controller (NanoScope V, Veeco, Plainview, USA) and a silicon tip (RTESP, Bruker, Billerica, USA) at a frequency of 318 kHz. For the sample preparation a droplet (3 µL) of highly diluted (ca 0.0001 wt%) microgels in water was dropped on a silicon wafer and dried overnight. The AFM measurements allowed the determination of the polydispersity in size of the dried microgels, needed as input for the Monte-Carlo simulations. The polydispersity is described with a Gaussian distribution function of the number of microgels per unit volume of a radius comprised between r and r+dr.
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4

Topography Analysis of HMP Variants

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The topography of the HMPcontrol, HMPIFUS582,864 and 1144 kHz -Aa(+) and HMPIFUS582,864 and 1144 kHz -Aa(−) was observed by Atomic Force Microscope (diMultiMode™V de Veeco Instruments - Nanoscope V, USA). The samples were placed on a freshly cleaved mica sheet and then air dried at room temperature. The HMP topography was evaluated from 3D surface plotting, height and phase analysis [36] .
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5

Wafer-Scale MoTe2 Layer Fabrication

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Wafer-scale 2D MoTe2 layers were fabricated via a thermal-assisted tellurization route. In detail, Mo precursor layers were first defined on pre-cleaned substrates by a magnetron sputtering system. Then, the Mo-coated substrates were placed on a quartz boat filled with Te powder (99.99%) at the center of a horizontal tube furnace. The tube was evacuated and filled with a gas mixture of Ar (95%) and H2 (5%) at 50 sccm. Afterward, the Mo samples and Te powder were heated to 700 °C. During the heating process, the MoTe2 layers can be produced by the transportation of vaporized Te to the Mo samples. The prepared 2D MoTe2 layers were analyzed by XRD (RigakuSmart Lab), Raman spectrometry (Horiba HR800), AFM (Veeco Nanoscope V), a field-emission-gun scanning electron microscope (LEO 1530) attached with an EBSD detector (Oxford Instruments NordlysNano EBSD detector and AZtecHKL), a UV/Vis/NIR spectrometer (PERKIN ELMER), STEM (JEOL JEM-ARM300F), and XPS (Thermo ESCALAB 250).
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6

Scaffold Morphology and Silk Microstructure

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The morphology of the scaffold samples was observed with SEM (Hitachi
S-4800, Hitachi, Tokyo, Japan). For SEM, the samples were imaged at 3.0 kV after
sputter-coated with gold.21 (link)The microstructures of silk in aqueous solutions were evaluated with AFM
(Nanoscope V, Veeco, NY, USA). 2 μL of the diluted samples (0.01
wt%) were dropped onto freshly cleaved mica surface and measured by AFM
with a spring constant of 3 N m−1 in tapping mode.
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7

Atomic Force Microscopy of DNA-Protein Complexes

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Sample solutions contained linear pCX eGFP DNA (by treatment with BcuI) at 10 ng/μl and/or the designated HU protein (3 μM) in buffer (10 mM HEPES, pH 7.5, 4 mM MgCl2). This protein: DNA ratio equals to the presence of 1 HU homodimer for every 10 bp DNA, if all proteins bind. The solution was either incubated at 37°C for 1 h or without incubation. Ten microliters of sample solution was applied to freshly cleaved mica by spin coating (15 s ramp 0–2,000 rpm; 30 s at 2,000 rpm), followed by washing with water (2 ml, full wetting with intervals at 3,000 rpm) and drying (4 min at 3,000 rpm).
Atomic force microscopy measurements were carried out in air at 293 K, using a Nanoscope V instrument (Veeco), type Multimode 8. Probes PPP-NCHR POINTPROBE-PLUS® Silicon-SPM-Sensor were used (resonance frequency = 330 kHz; force constant = 42 N/m, length = 125 μm, NanosensorsTM) operating in tapping mode at a frequency around 321.5 kHz to image the surfaces through ScanAsyst®-air mode. The data were processed using the WS × M software (Horcas et al., 2007 (link)).
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8

Thin-Film Surface Characterization via AFM

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Advanced imaging modes have
been developed via AFM characterization using MultiMode scanning probe
microscope (Nanoscope V, Veeco, Santa Barbara) in tapping mode to
examine surface morphologies. The thin-film thickness was measured
using a surface profilometer (Bruker’s Dektak 150, Veeco, New
York).
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9

Atomic Force Microscopy of Mycobacterial Cell Surfaces

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An atomic force microscope was used to visualize the surface of the mycobacterial cells after the treatment with the S. hermaphrodita seed extract. The analysis was performed using a NanoScope V atomic force microscope (Veeco Instruments Inc., Santa Barbara, CA, USA) equipped with a scanning probe. The measurements were made in the PeakForce QNM mode using the RTESPA probe. Three 2 µm × 2 µm fields were probed in each sample. The roughness was read from 10 fields with dimensions of 100 nm × 100 nm from each 2 μm × 2 μm image, and then the arithmetic mean was calculated from the results. Nanoscope Analysis version 1.4 (Veeco, USA) and WSxM version 5.0 (Nanotec, Madrid, Spain) programs were used to analyze the images.
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10

Visualizing Peptide-pNIPAm Hybrid Nanostructures

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The morphology
and size of the peptide-pNIPAm SAPEs were visualized by transmission
electron microscopy (TEM). Briefly, samples (0.1 mg/mL in HEPES 10
mM) were soaked onto prewarmed carbon-coated copper grids for 2 min,
and excess liquid was removed by a filter paper. The grids were subjected
to negative staining with 2% uranyl acetate (Merck, Germany) for 45
s and dried for 10 min at room temperature before acquisition of TEM
images (Tecnai 10, Philips, The Netherlands). Atomic force microscopy
(AFM) was applied to visualize the morphology of the peptide-based
vaccines. Briefly, 40 μL of a 0.02% (w/v) polylysine solution
in water was placed on a freshly cleaved mica substrate and, after
5 min incubation, washed with filtered Milli-Q water followed by drying
under N2 flow. Next, 30 μL of the sample was loaded
on the treated mica and incubated for 1 min, followed by washing three
times with 20 μL of filtrated Milli-Q water to discharge nonattached
particles and salts. Finally, the prepared sample was dried by an
N2 flow. ScanAsyst image mode was used at room temperature
with a Digital Instruments NanoScope V, equipped with silicon nitride
cantilevers (Veeco, NY, USA). Images were analyzed by NanoScope Analysis
1.40 software. Several images were taken at different days of individually
prepared samples.
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