AFM imaging was performed to characterize the microstructure of the different hydrogels at the fiber level. A 100 μl sample from each hydrogel composition (n = 3) was deposited on a silica sample holder and incubated at 37 °C for 30 min. After gelation, the hydrogel's top surface was carefully removed using gentle air blowing/drying to allow the imaging of the internal network. Agilent 5500 with MAC III controller and JPK Nanowizard 3 AFM systems were used for morphological imaging in intermittent contact mode in air. A sharp microlever probe MSNL-F (f = 120 kHz, k = 0.6 N/m, tip radius of curvature < 12 nm) was used. The AFM topography images were leveled, line-corrected, and measured (height and roughness profiles) using Gwyddion48 (link). A fiber extraction algorithm, ct-FIRE49 (link), was applied to the AFM images to characterize the fiber diameter, length, straightness, and alignment.
Jpk nanowizard 3
Manufactured by Bruker
Sourced in Germany, United States
The JPK NanoWizard 3 is an atomic force microscope (AFM) system designed for high-resolution imaging and analysis of samples at the nanoscale level. It provides a versatile platform for studying the topography, mechanical properties, and other characteristics of a wide range of materials, including polymers, biomaterials, and small-scale electronic components.
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Lab products found in correlation
Axio observer z1, by Zeiss (9 mentions)
Matlab, by MathWorks (2 mentions)
Hq nsc18 al bs cantilevers, by MikroMasch (2 mentions)
Agilent 5500, by Agilent Technologies (1 mentions)
Ix81 s1f 3, by Olympus (1 mentions)
X pert system, by Philips (1 mentions)
Cary 5000 spectrophotometer, by Agilent Technologies (1 mentions)
Petri dish heater, by Bruker (1 mentions)
Data processing software, by Bruker (1 mentions)
Hq nsc15 al bs cantilevers, by MikroMasch (1 mentions)
Scanasyst fluid, by Bruker (1 mentions)
Cell tak, by Corning (1 mentions)
Afm probe, by Bruker (1 mentions)
Prism 5, by GraphPad (1 mentions)
Pnp tr 20, by NanoWorld (1 mentions)
Arrow tl1 cantilevers, by NanoWorld (1 mentions)
Dnp s, by Bruker (1 mentions)
Dimension icon, by Bruker (1 mentions)
Scanasyst air cantilever, by Bruker (1 mentions)
Omcl ac160ts, by Olympus (1 mentions)
28 protocols using jpk nanowizard 3
1
Characterizing Hydrogel Fiber Microstructure
2
Measuring Cardiomyocyte Contractility with AFM
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Atomic force microscope (JPK NanoWizard 3, JPK, Berlin, Germany), combined with an inverted light microscope (IX-81S1F-3, Olympus, Tokyo, Japan), was used to obtain mechanocardiograms of beating EBs as previously described (Pesl et al., 2014 (link); Caluori et al., 2019 (link); Pribyl et al., 2019 (link)). Drug response tests were performed after initial equilibration in Tyrode's solution, followed by addition of 70 μM metoprolol, 10 min of stabilization time, and then 10 min of measurement. Isoproterenol (1 μM) was subsequently added into the solution, and mechanocardiograms were measured after 20 min of stabilization. The mechanical properties of beating EBs were expressed as force of contraction in nN as previously described (Pesl et al., 2014 (link)); however, the values were relativized for final comparison.
3
Atomic Force Microscopy of Hydrogel Fibers
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Freshly
cleaved mica sheets were rinsed with deionized water and dried under
a nitrogen flow. For the pregelation samples, a drop of the viscous
solution (10 mM) was deposited on the mica sheet. For gel samples,
a mica sheet was placed on the top of the gel (10 mM) for 1 min, washed
with water, and dried under a gentle stream of nitrogen. AFM images
were acquired using JPK Nanowizard 3 (JPK, Germany) in air at room
temperature under AC mode. The scans were acquired at a 512 ×
512 pixel resolution using ACTA-SS probes (k ∼37
N/m, AppNano, Scientec, France), a drive frequency of ∼254
kHz, a set point of ∼0.5 V, and a scanning speed of 1.0 Hz.
Images were analyzed using the JPK data processing software. The fiber
diameter was determined from the generated AFM images using the ImageJ
software.
cleaved mica sheets were rinsed with deionized water and dried under
a nitrogen flow. For the pregelation samples, a drop of the viscous
solution (10 mM) was deposited on the mica sheet. For gel samples,
a mica sheet was placed on the top of the gel (10 mM) for 1 min, washed
with water, and dried under a gentle stream of nitrogen. AFM images
were acquired using JPK Nanowizard 3 (JPK, Germany) in air at room
temperature under AC mode. The scans were acquired at a 512 ×
512 pixel resolution using ACTA-SS probes (k ∼37
N/m, AppNano, Scientec, France), a drive frequency of ∼254
kHz, a set point of ∼0.5 V, and a scanning speed of 1.0 Hz.
Images were analyzed using the JPK data processing software. The fiber
diameter was determined from the generated AFM images using the ImageJ
software.
4
Topography Imaging of Gel Structures
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The gels were placed on clean glass coverslips and dried under vacuum. Topography images of the structures were taken using a JPK NanoWizard®3 (JPK Instruments, Berlin, Germany) working in AC mode. Si3N4 cantilever probes with a nominal spring constant of 3 Nm−1 and a resonance frequency of 75 kHz were used.
5
Atomic Force Microscopy Topographic Imaging
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Atomic force microscopy (AFM) topographic imaging was employed to measure the roughness of the samples. Imaging was performed on a JPK NanoWizard 3 (JPK Instruments AG) operated in ac mode using ACTA cantilevers (spring constant ≈40 N/m, resonance frequency ≈300 kHz; Applied NanoStructuresInc.). Image processing was carried out with JPK SPM Data Processing.
6
Force Spectroscopy of Lipid Bilayers
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Force spectroscopy measurements were carried in aqueous solution at RT to determine the forces of lipid bilayers in PBS buffer using JPK NanoWizard 3 (JPK Instruments AG, Berlin, Germany). Cantilevers (OBL-10, nominal spring constant of ∼6 pN nm−1 and nominal tip radius of 30 nm from Bruker, Ettlingen, Germany) were UV-ozone treated (Pro Cleaner Plus; BioForce Nanoscience, Virginia Beach, VA) for 30 min and calibrated using thermal method. The calibration was first done in air against mica and then in PBS buffer. The calibration software provided in the JPK instrument recorded the new spring constant and deflection sensitivity that was used further. A deviation of 10–20% from the company mentioned spring constant was observed. The force curves obtained (approach speed of 1 μm/s) were processed using JPK Data Processing software (version 5.0.91) and analyzed using a home-written MATLAB (The MathWorks, Natick, MA) script. To determine the peak adhesive forces, a histogram of the forces obtained was plotted and Kernel density estimation (KDE) method (nonparametric method for multivariate distribution analysis) was applied (with Gaussian kernel) to determine the peak position.
7
Nanomaterial Characterization Techniques
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Transmission electron microscopy (TEM) images and selected area electron diffraction (SAED) patterns were obtained on a JEOL-1011 (100 kV). Samples for the TEM analysis were prepared by drop-casting 10 μL dispersion of nanocrystals diluted in toluene onto carbon-coated copper or titanium TEM grids followed by solvent evaporation. High resolution (HR) TEM images were obtained with a Philips CM 300 UT microscope operated at 200 kV. X-ray diffraction (XRD) measurements were performed with a Philips X'Pert System with Bragg–Brentano geometry, equipped with a copper anode (Kα X-ray wavelength of 0.154 nm). Samples were prepared by drop-casting the colloidal nanocrystal solution onto silicon wafer substrates (<911> or <711> cut) with subsequent solvent evaporation. Atomic force microscopy (AFM) measurements were performed on a JPK Instruments system (JPK Nano Wizard 3) in intermittent contact mode. UV-VIS-NIR absorption spectra were obtained with a Cary 5000 spectrophotometer equipped with an integration sphere.
8
Microsphere Mechanical Characterization by AFM
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Mechanical characterization of microspheres was measured by AFM on a JPK Nanowizard 3 (JPK instruments, Germany) coupled with an inverted microscope (NIKON Ti, Nikon Instruments) keeping a constant temperature through a Petri Dish Heater (JPK instruments, Germany).
9
Probing Polymerized Proptose Emulsions
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Mechanical properties of the polymerized proptose emulsions were performed in aqueous suspension using an AFM (JPK NanoWizard 3 from JPK, Berlin, Germany) equipped with qp‐BioAC CB1 cantilever (uniqprobe, NanoWorld AG) with a spring constant of 0.2 N m−1, 7 µm tip height, and 8 nm tip radius. Sample preparation included a coating of the glass coverslip with a positively charged poly‐l‐ornithine solution for 20 min, followed by washing with Milli‐Q water. Particle suspension was allowed to settle down for 15 min before scanning. 3D‐scanner with 15 µm piezo‐driven stroke in the z‐direction and 100 µm in the x‐ and y‐direction was used. 5 × 5 µm size was scanned with different set points at 1, 5, 10, 15, and 20 nN (for the block copolymer membranes) and 1, 2, 3, and 4 nN (for the polymerized zone). Data processing was performed using JPK Data processing software.
10
Topographic Analysis of Surfaces by AFM
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The topographies of all surfaces were analyzed by atomic force microscopy (AFM) using a JPK NanoWizard 3 (JPK GmbH, Berlin, Germany) and HQ:NSC15/AlBS cantilevers (MikroMasch, Wetzlar, Germany) in intermittent contact mode in air. The images had a size of 5 × 5 µm 2 and a resolution of 1024 × 1024 px. All AFM images were processed using Gwyddion (version 2.51) [22] (link) and the RMS surface roughness Sq was obtained for each image. The Sq values were averaged over four images and the standard deviations are given as errors.
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