MVs were isolated from differentiating chondrocytes by differential centrifugation as described earlier.(9 (link)) A drop (5 µL) of each MV solution in Tris-buffered-saline was spotted on freshly cleaved mica substrates (Ted Pella, Redding, CA) and allowed to stand for 5 min. Next, 5 µL of glutaraldehyde solution (8% in H2O, Sigma-Aldrich, St. Louis, MO) was dropped onto the samples. The substrates were stored inside a desiccators at room temperature for 24 h. AFM images of dried samples were recorded in air by means of an 5500 atomic force microscope (Agilent Technologies, Santa Clara, CA) equipped with an open-loop probe working in non-contact (AAC) mode. Silicon-nitride cantilevers having a nominal resonance frequency of ~190 kHz (NanosensorsTM, Neuchatel, Switzerland) were used. Tridimensional AFM images were generated by PicoView software (Agilent Technologies). AFM images were used to gather information about the morphology, height, volume and number of MVs in each sample. AFM phase images were also recorded on samples prepared without the use of glutaraldehyde.
5500 atomic force microscope
Manufactured by Agilent Technologies
Sourced in United States
The 5500 atomic force microscope is a versatile instrument designed for high-resolution imaging and analysis of surface topography. It utilizes a sharp probe to scan and measure the surface of a sample, providing detailed information about its physical characteristics at the nanoscale level.
Automatically generated - may contain errors
Lab products found in correlation
Mica substrate, by Ted Pella (1 mentions)
Glutaraldehyde solution, by Merck Group (1 mentions)
Picoview software, by Agilent Technologies (1 mentions)
Jem 2100f, by JEOL (1 mentions)
Uv 3600 spectrophotometer, by Shimadzu (1 mentions)
Vetex70, by Bruker (1 mentions)
Zetasizer zs90, by Malvern Panalytical (1 mentions)
Uv 3600, by Shimadzu (1 mentions)
Bi 200sm, by Brookhaven Instruments (1 mentions)
Jem 2100, by JEOL (1 mentions)
Ppp zeilr, by Nanosensors (1 mentions)
14 protocols using 5500 atomic force microscope
1
Atomic Force Microscopy of Chondrocyte-Derived Extracellular Vesicles
2
Comprehensive Characterization of Gold Nanoplates
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The morphologies of the gold nanoplates were characterized using a JEOL200CX transmission electron microscope (TEM) operated at 200 kV. Selected area (electron) diffraction (SAED) and HRTEM images were obtained on a JEM-2100F (JEOL Ltd., Japan). Scanning electron microscopy (SEM) images were obtained from an S-3000 N scanning electron microscope (Tokyo, Japan). Atomic force microscopy (AFM) images were recorded on a 5500 Atomic Force Microscope (Agilent Technologies, USA). UV-vis absorption spectra were recorded on a SHIMADZU UV-3600 spectrophotometer (Japan). The scattering spectrum was obtained with a monochromator (Acton SP2358, PI, USA) (grating density: 300 lines per mm; blazed wavelength: 500 nm) combined with liquid nitrogen cooled PyLoN ccd cameras (Princeton Instruments, USA). Traditional electrochemical measurements were performed using a CHI-630A (CH Instruments Co.) with a three-electrode system at room temperature.
3
Characterization of MoS2 Nanostructures
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The morphology of MoS2 was measured using transmission electron microscopy (TEM; JEM-2100, JEOL). The thickness and size of the MoS2 particles were determined with a 5500 atomic force microscope (AFM; Agilent). The zeta potential was quantified with a ZS90 Zetasizer instrument (Malvern Instruments). Dynamic light scattering (DLS) was performed with static light scattering instrument (BI-200SM, Brookhaven Instruments). UV–vis spectra were obtained on a UV3600 instrument (Shimadzu Corporation). Fourier transform infrared (FT-IR) spectroscopy was recorded on a Vetex70 (Bruker Corp., Germany). The photothermal properties of the composite were examined using a laser device (Shanghai Xilong Optoelectronics Technology Co. Ltd.) at a wavelength of 808 nm.
4
Characterization of Polysaccharide Nanoparticles
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PZMP3-1 used the XRD pattern to determine the crystal structure present. The angular range of the diffractometer was 5–50° (2θ), the step size was 0.01°, the scan speed was 15°/min, and tube pressure (40 kV) and tube flow (40 mA) were present. Using an SEM (S-4800, Japan), the morphological characteristics of PZMP3-1 were documented. We used the Cressington 208 HR Sputtering Coater with sputtering gold samples. We dissolved the polysaccharides with distilled water and dried the test samples dripping on the mica carrier surface at ambient pressure of 70°C. A 5500 atomic force microscope (Agilent) was used to create the AFM images (30 (link)).
5
Nanoscale Biomechanical Characterization of Cells
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The morphology and biomechanical properties of the YSE2-FAT1shRNA and Colo680N-FAT1shRNA cells and the corresponding controls were detected by AFM, as previously described (19 (link)). Briefly, AFM images were acquired in the tapping mode and in contact mode using Si3N4 tips (NSC19, 0.68 N/m normal spring constant; Schaefer Technologie GmbH, Langen, Germany) and gold-coated tips (CSC 38; Schaefer Technologie GmbH), respectively. AFM measurements were performed in aqueous solution at room temperature, using a 5500 atomic force microscope (Agilent Technologies, Inc., Santa Clara, CA, USA). The spring constants of the cantilevers used for AFM force spectroscopy were ~0.1 N/m. Adhesion and elasticity maps were obtained by recording 16×16 force-distance curves on areas of a given size (2×2 µm), calculating the adhesion force and elasticity modulus for each force curve and displaying these values as gray and colorized scale pixels, respectively. These maps qualitatively and quantitatively demonstrated the viscoelasticity of individual cells at the nanoscale level.
6
Atomic Force Microscopy Surface Roughness Measurement
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Atomic force
microscopy (AFM) measurements were performed with a 5500 Atomic Force
Microscope (Agilent Technologies) using an arrowshaped cantilever
(PointProbe Plus ZEISS Veritekt Microscopes - Contact Mode Low Force
Constant - Reflex Coating (PPP-ZEILR), Nanosensors, tip diameter <
10 nm). All images were recorded in the intermittent contact mode
with constant force. The experiments were performed in a glovebox
with argon flow to minimize contact to air. An area of 5 μm
× 5 μm was chosen for all measurements. For data processing,
the software MountainsSPIP (Digital Surf/Image Metrology) was utilized.
The calculation of the arithmetic mean deviation of the surface roughness
(called average surface roughness, Sa,
for simplicity) was done according to EUR 15178N. The maximal surface
roughness (Sm) is the difference between
the highest and the lowest point on the sample surface within the
region of interest.
microscopy (AFM) measurements were performed with a 5500 Atomic Force
Microscope (Agilent Technologies) using an arrowshaped cantilever
(PointProbe Plus ZEISS Veritekt Microscopes - Contact Mode Low Force
Constant - Reflex Coating (PPP-ZEILR), Nanosensors, tip diameter <
10 nm). All images were recorded in the intermittent contact mode
with constant force. The experiments were performed in a glovebox
with argon flow to minimize contact to air. An area of 5 μm
× 5 μm was chosen for all measurements. For data processing,
the software MountainsSPIP (Digital Surf/Image Metrology) was utilized.
The calculation of the arithmetic mean deviation of the surface roughness
(called average surface roughness, Sa,
for simplicity) was done according to EUR 15178N. The maximal surface
roughness (Sm) is the difference between
the highest and the lowest point on the sample surface within the
region of interest.
7
Nanomaterial Surface Characterization
8
Characterizing Film Topography and Structure
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The surface topography of the films was measured with an Agilent 5500 Atomic Force Microscope operated in the acoustic mode. GIWAXS patterns were collected at the SAXS/WAXS beamline at the Australian Synchrotron68 using a Pilatus 1 M photon detector. Photons (9 keV) were aligned parallel to the surface of the samples using a crystal analyser. Scattering patterns were collected at incident angles ranging from 0.05 to 0.25°, with the images reported taken at the critical angle identified by the angle with the highest scattering intensity. NEXAFS spectra were collected at the SXR beamline of the Australian Synchrotron69 and analysed using previously used protocols44 (link).
9
Fabrication of Nanocrystal Thin Films
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For nanocrystal thin films, the
colloidal solutions are purified once more with ethanol, centrifugation,
and redissolving in chloroform. Filtered solutions are dropped on
clean substrates and spin coated to form a single monolayer (approx.
15 nm thickness). Films are soaked for 30 s with TBAI solution in
methanol (20 mg/mL) to remove surface ligands and subsequently spin-washed
three times with pure methanol. For thicker films, the deposition
cycle is repeated on top of a previous layer. The thickness is measured
with an Agilent 5500 Atomic force microscope in tapping mode.
colloidal solutions are purified once more with ethanol, centrifugation,
and redissolving in chloroform. Filtered solutions are dropped on
clean substrates and spin coated to form a single monolayer (approx.
15 nm thickness). Films are soaked for 30 s with TBAI solution in
methanol (20 mg/mL) to remove surface ligands and subsequently spin-washed
three times with pure methanol. For thicker films, the deposition
cycle is repeated on top of a previous layer. The thickness is measured
with an Agilent 5500 Atomic force microscope in tapping mode.
10
Atomic Force Microscopy Characterization
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Atomic force microscopy study was carried out in tapping mode using an Agilent 5500 atomic force microscope in ambient atmosphere. The approximate resonance frequency of the cantilever was 280 kHz and force constant was ≈60 Nm−1.
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