AFM images were collected for characterizing morphology of scratch and scratch-etched Si wafer. NanoScope III microscope (Digital Instruments, Inc., Santa Barbara, CA) and silicon cantilevers TAP300Al-G (Ted Pella, INC, Redding, CA) having force constant 40 N/m and resonance frequency of 300 kHz were used for imaging in tapping mode. The images were processed using Gwyddion software.
Tap300al g
Manufactured by Ted Pella
Sourced in United States
The TAP300Al-G is a benchtop scanning electron microscope (SEM) designed for high-resolution imaging and analysis of samples. It features a tungsten electron source, a maximum accelerating voltage of 30 kV, and can achieve a resolution of up to 5 nm. The instrument is equipped with a 300 mm X-Y motorized stage and can accommodate a variety of sample types.
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5 protocols using tap300al g
1
Characterization of Scratch-etched Silicon Wafers
2
Atomic Force Microscopy Surface Scratching
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The developed surfaces were scanned by AFM (Cypher, Asylum Research) in air tapping mode using a cantilever with a tip radius <10 nm (Tap300AL-G, TED PELLA, INC). For sample scratching, we used the same tip in contact mode, and the force applied to the tip (F) was controlled by the applied voltage (U) with a calibrated correlation of F(nN) = 600U(V). To create line scratches, the tip was moved at a speed of 1 μm/second. Area scratches were made by scanning the area (200 nm × 200 nm) of interest with a resolution of 256 × 256 at contact mode at a frequency of 1 Hz.
3
Characterizing Surface Topography and Protein Coatings
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Master and replica surfaces were imaged using a Hitachi S-4700 SEM. The surfaces were sputter-coated with Pt/Pd in argon plasma at 20 mA for 25 s (Cressington sputter coater 108). The height/depth of the structures was characterized by a Veeco Multimode AFM in tapping mode using aluminum-coated silicon probes (TAP300AL-G, Ted Pella).
A collagen-coated surface treated with Alexa Fluor 594 NHS ester was imaged in PBS using a Leica SP5 X confocal microscope with a 100× objective (NA 1.4) and a scanner zoom factor of 4. Images were obtained in a 512 × 512 pixel format. The fluorescent-fibronectin-coated surface was imaged in HBSS in the same manner. The orthogonal view is a single slice reconstructed from z-stack images.
A collagen-coated surface treated with Alexa Fluor 594 NHS ester was imaged in PBS using a Leica SP5 X confocal microscope with a 100× objective (NA 1.4) and a scanner zoom factor of 4. Images were obtained in a 512 × 512 pixel format. The fluorescent-fibronectin-coated surface was imaged in HBSS in the same manner. The orthogonal view is a single slice reconstructed from z-stack images.
4
Characterizing Morphology of np-Au via AFM
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Atomic force microscopy (AFM) images were collected in tapping mode for characterizing the morphology of np-Au before and after DOX loading, and the corresponding height distribution profiles were plotted to determine the changes in height. BioScope ResolveTM AFM (Bruker, Santa Barbara, CA, USA) integrated with NanoScope® software and silicon cantilevers TAP300Al-G (Ted Pella, INC, Redding, CA, USA) with a force constant of 40 N/m were used for imaging. The images were processed using Gwyddion software.
5
AFM Nanoindentation of Phage Fibers
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The Asylum MFP-3D (Asylum Research, Santa Barbara, CA), Igor Pro 6.0, and Asylum software package was used to measure the mechanical properties of electrospun phage fibers through AFM nanoindentation method. In this method, the probe was a cone shaped silicon tip (Tap300Al-G, Ted Pella, Inc., Redding, CA) with <10 nm radius, and 62 N m−1 spring constant. The force-indentation loading and unloading curves of 50 different points in several phage electrospun fibers were taken with AFM nanoindentation method. According to the Oliver–Pharr model,41 (link) a line with slope S = dF dl−1 (S: unloading stiffness, F: force, l: indentation length) was fit in the upper portion of the unloading curves in the data to calculate Young's modulus.42 (link) We used dehydrated electrospun phage fibers on the gold-coated silicon substrate under environmental conditions (25 °C, 40% relative humidity).
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