All measurements were conducted using a commercial atomic force microscope (NTEGRA AFM, NT-MDT) and V-shaped silicon nitride AFM probes (PNP-TR-Au, Nanoworld) with a spring constant of 0.08 N/m and face angle of 35° at room temperature. The cantilever spring was calibrated by thermal noise fluctuation methods,46 and the deflection sensitivity of each probe was calibrated by force-distance measurements on the bare glass area of the petri dish. For cell experiments, all force-distance curves were obtained from the central cytoplasmic region of the cell surface. At least 256 force-distance measurements were carried out per individual cell. The loading rates ranging from 235 – 6720 pN/s were calculated by multiplying the probe retracting rate by the effective spring constant of the cantilever-cell membrane system. A 50 μg/ml of free cRGD peptide solution was used for integrin blocking experiments. The scanning resolution was 256 × 256 pixels with a scan rate of 0.1 – 0.5 Hz, depending on the scanning areas of irregular cell size. The acquired images were flattened, if required, to eliminate the background noise and tilt from the surface using all unmasked portions of scan lines to calculate individual least-square fit polynomials for each line.
Ntegra afm
Manufactured by NT-MDT
The NTEGRA AFM is a versatile atomic force microscope (AFM) system designed for advanced surface characterization. It provides high-resolution imaging and analysis capabilities for a wide range of applications.
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6 protocols using ntegra afm
1
Quantifying Cell Membrane Mechanics
2
Multimodal Nanoscale Imaging of Decorated Nanotubes
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Tilt series of high-resolution
STEM HAADF images were acquired on an FEI F20 TWIN electron microscope.
The tilt increment was 2° in a range of 132°. Tomograms
were reconstructed using the iterative simultaneous reconstruction
technique, after cross-correlation alignment and iterative feedback
displacement refinement.41 (link),42 (link) MeVisLab was used for
the visualization of tomogram data.
AFM measurements were performed
using an NTEGRA AFM with Smena head (NT-MDT, Zelenograd). A module
for performing scanning thermal microscopy—VertiSense (App
Nano, Mt. View, CA) was applied to investigate the thermal properties
of decorated and uncoated nanotubes. VTP-200 probes, with nominal
spring constant of 10 N/m, were used in the semicontact mode. The
module was operated in the heat dissipation mode, where the laser
alignment on the probe was adjusted to heat the tip to approximately
20 K above room temperature, so that mappings of heat dissipation
could be obtained simultaneously with the topography. SEM micrographs
were made of the sample before and after imaging the nanotubes by
AFM.
STEM HAADF images were acquired on an FEI F20 TWIN electron microscope.
The tilt increment was 2° in a range of 132°. Tomograms
were reconstructed using the iterative simultaneous reconstruction
technique, after cross-correlation alignment and iterative feedback
displacement refinement.41 (link),42 (link) MeVisLab was used for
the visualization of tomogram data.
AFM measurements were performed
using an NTEGRA AFM with Smena head (NT-MDT, Zelenograd). A module
for performing scanning thermal microscopy—VertiSense (App
Nano, Mt. View, CA) was applied to investigate the thermal properties
of decorated and uncoated nanotubes. VTP-200 probes, with nominal
spring constant of 10 N/m, were used in the semicontact mode. The
module was operated in the heat dissipation mode, where the laser
alignment on the probe was adjusted to heat the tip to approximately
20 K above room temperature, so that mappings of heat dissipation
could be obtained simultaneously with the topography. SEM micrographs
were made of the sample before and after imaging the nanotubes by
AFM.
3
AFM Characterization of Surface Morphology
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AFM was performed with an NTEGRA AFM (NT-MDT Spectrum Instruments, Moscow, Russia) in semi-contact mode (tapping mode) and Nova Px 3.5.0 software. NGS01 tips from NT-MDT Spectrum Instruments with a typical tip radius of 6 nm were used. The scan parameters were optimized using the ScanT software extension in the attractive measurement regime. Scans were performed over an area of 10 × 10 µm2 and a resolution of 512 pixels × 512 pixels for each sample. Each measurement line was recorded in two measuring directions. Based on the two images, a minimum was calculated, minimizing the parachuting effect. Subsequently, the images were aligned using a first-order line fit. Surface roughness was calculated using the integrated roughness analysis over the whole surface of the 10 × 10 µm2 scans. The peaks were cut in some images to allow a better comparison of structures.
4
High-speed AFM of PNIPAM Thermal Response
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High-speed AFM was performed as described previously24 (link). Drift correction was performed to keep the pore centered. The temperature was increased by heating the whole measurement setup. A thermometer was used to verify that the solution has crossed the LCST of PNIPAM.
Contact mode images were obtained in water using an NTEGRA AFM (NT-MDT) with Tap300AI-G tips (BudgetSensors). In Gwyddion (http://gwyddion.net/ ) mean plane subtraction was used to level the data, as well as row alignment with a 2nd degree polynomial.
Contact mode images were obtained in water using an NTEGRA AFM (NT-MDT) with Tap300AI-G tips (BudgetSensors). In Gwyddion (
5
Characterization of Polymersomes by DLS and AFM
6
Characterization of Engineered T4L Mutants
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