Atomic Force Microscopy (AFM) observations were carried out with the “J scanner” in tapping mode by using a Nanoscope IIIA-MultiMode AFM (Digital Instruments, Santa Barbara, CA, USA) under room conditions. The force was maintained at the lowest possible value by a continuous adjusting of the set point during the imaging phase. Images were recorded using 0.5–2 Ω·cm phosphorous (n) doped silicon tips mounted on cantilevers with a nominal force constant of 40 N/m, a resonance frequency of 300 kHz and a tip curvature radius of 10 nm.
Nanoscope iiia multimode afm
Manufactured by Digital Instruments
Sourced in United States
The Nanoscope IIIa Multimode AFM is a high-resolution scanning probe microscope designed for topographic and nanoscale characterization of surfaces. It utilizes atomic force microscopy (AFM) technology to provide detailed imaging and analysis of samples at the nanometer scale. The core function of this instrument is to enable users to acquire precise, high-quality surface data without interpretation or extrapolation.
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7 protocols using nanoscope iiia multimode afm
1
Atomic Force Microscopy Imaging Procedure
2
Characterizing NP Surface Morphology by AFM
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NP surface morphology was characterized by AFM, using a Nanoscope IIIa Multimode AFM (Digital Instruments, Veeco). Samples were prepared by depositing a drop of final colloidal suspension (10 mg mL−1) onto freshly cleaved mica for 15 min at room temperature and dried with pure nitrogen. Samples were analyzed in tapping mode in air at room temperature using etched silicon tips (≈300 kHz), at a scan rate of ≈1.6 Hz.
3
Atomic Mica Surface Functionalization for Viral Imaging
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An atomically flat mica (grade V-I, SPI) surface functionalized with positively charged poly-L-lysine (Sigma Aldrich) was chosen to assure the attachment of viral particles for AFM imaging: 10 µl poly- L-lysine (0,01%) were deposited onto freshly cleaved mica-discs. After an incubation of 30 seconds, disks were rinsed twice with 1 ml of ultrapure water and dried by a nitrogen flow. A 5 µl-droplet of the viral agent obtained after purification was applied to the surface and incubated for two minutes (VLPs) or 15 minutes (viral cores) to assure a sufficient adsorption.
Imaging was performed in Tapping Mode with a Nanoscope IIIa Multimode AFM operated with a JVH- scanner (Digital Instruments Veeco, Santa Barbara). Since our main objective in this work is to compare size distributions of VLPs under different production conditions, we chose to image dried samples, although this might flatten particles, in order to avoid desorption of particles during scanning of the surface by the AFM tip. For imaging in air, the sample was rinsed with 1 ml of ultrapure water and gently dried by a nitrogen flow. Imaging was executed with silicon tips (resonance frequency ∼350 kHz, kcantilever = 40 N/m). According to the scan size of 0.5–3 µm, a scan rate between 1 and 3 Hz was chosen.
Additional AFM imaging in liquid conditions are provided as supplemental data inFile S1 .
Imaging was performed in Tapping Mode with a Nanoscope IIIa Multimode AFM operated with a JVH- scanner (Digital Instruments Veeco, Santa Barbara). Since our main objective in this work is to compare size distributions of VLPs under different production conditions, we chose to image dried samples, although this might flatten particles, in order to avoid desorption of particles during scanning of the surface by the AFM tip. For imaging in air, the sample was rinsed with 1 ml of ultrapure water and gently dried by a nitrogen flow. Imaging was executed with silicon tips (resonance frequency ∼350 kHz, kcantilever = 40 N/m). According to the scan size of 0.5–3 µm, a scan rate between 1 and 3 Hz was chosen.
Additional AFM imaging in liquid conditions are provided as supplemental data in
4
Atomic Force Microscopy of G-Wire Structures
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Atomic force microscopy (AFM) imaging was performed in tapping mode using a Nanoscope IIIa-MultiMode AFM (Digital Instruments, Santa Barbara, CA) equipped with the E (10 μm) scanner. We used silicon cantilevers (Bruker OTESPA-R3), with nominal resonance frequency 300 kHz and nominal tip radius of 7 nm.
Height and length distribution where obtained with ImageJ and the Ridge Detection plugin (https://imagej.net/Ridge_Detection ) based on the algorithm for detecting ridges and lines described by Steger47 (link)–49 . The height and length histograms for d(G2AG4AG2) G-wires were obtained by detecting all G-wires (N = 907) in eight AFM images (Supplementary Fig. 1 ). The height and length histograms for d(G2AG4C G2) G-wires were obtained by taking into account all detected G-wires (N = 3766) in 12 AFM images (Supplementary Fig. 24a ). In case of (G2AG4C G2) G-wires deposited on mica, which was not pre-treated with saturated solution of MgCl2, the height and length histograms were obtained by taking into account all detected G-wires (N = 2646) in 20 AFM images (Supplementary Fig. 24b ). One should note that the actual G-wire lengths are around 10 nm shorter than what is observed by AFM due to the finite tip size effect50 .
Height and length distribution where obtained with ImageJ and the Ridge Detection plugin (
5
Atomic Force Microscopy of Materials
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AFM measurements were carried out with the “J scanner” in tapping mode by using a Nanoscope IIIA-MultiMode AFM (Digital Instruments, Santa Barbara, CA, USA) under room conditions. The force was maintained at the lowest value as possible by continuous adjusting the set point during imaging. Images were recorded using 0.5–2 Ω·cm phosphorous (n) doped silicon tips mounted on cantilevers with a nominal force constant of 40 N/m, a resonance frequency of 300 kHz and a tip curvature radius of 10 nm.
6
Antibacterial Efficacy of Compound 1 on B. cereus
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Samples containing B. cereus ATCC 14579 (106 CFU mL−1) in MH medium were incubated with compound 1 at various concentrations (0, 8, and 16 µg mL−1) and after 2 h aliquots were taken. A drop (40 µL) containing the bacteria was deposited onto freshly cleaved mica surfaces for 30 min, gently washed with Milli-Q water, and dried under mild nitrogen flux. The surface was examined ex situ, at ambient temperature (21 °C), using Nanoscope IIIa multimode AFM (Digital Instruments, Veeco, Santa Barbara, CA) and etched silicon tips (TESP-V2, Bruker) with a resonance frequency of ca. 300 kHz. Images were acquired with scan rates between 1.2 Hz and 1.5 Hz. It is worth to note that washing and drying steps, as well as tip repetitive scanning did not influence the results presented in this work.
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Nanoscale Characterization of Nanoparticles
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