A Dimension Fastscan with ScanAsyst™ AFM (Bruker, Santa Barbara, CA) was used to analyzed biophysical changes in a single living cell [31 (link)-35 (link)]. The pyramidal tip of silicon nitride cantilevers (nominal spring constant k=0.01N/m, tip radius 20.0nm, tip half angle 18.0°, MLCT-C, Bruker, USA) was used for the contact mode of AFM. This experiment was performed as described previously [35 (link)]. The cells were imaged over an area of 20×20μm2 with a resolution of 128×128 pixels. NanoScope Analysis (Bruker, USA) was used to calculate cell elasticity values and roughness values. The Young's modulus was calculated using Hertz model, which was selected from 10-12loci (2μm×2μm) on each cell [33 (link)].
Scanasyst afm
Manufactured by Bruker
Sourced in Germany
The ScanAsyst AFM is an atomic force microscope (AFM) developed by Bruker. It is designed to provide high-resolution imaging and advanced analytical capabilities for a variety of samples. The ScanAsyst AFM utilizes innovative technology to automate and simplify the imaging process, making it accessible to users with varying levels of expertise.
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3 protocols using scanasyst afm
1
Biophysical Characterization of Living Cells
2
Polysaccharide Morphology by AFM
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The molecular morphology of polysaccharides was recorded using a ScanAsyst AFM (Bruker, Karlsruhe, Germany).
3
Tau Fibril Characterization on Mica
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Samples were prepared by splitting Mica discs (#50, 9.9-mm diameter; Ted Pella, Inc, PELCO Mica discs) with a fresh razor and sticking them to a glass coverslip with a double-sided sticky tab (#16084-6, 6 mm OD; Ted Pella, Inc, PELCO Tabs). Tau extracts were diluted to a tau concentration of 5 ng/μl, and Mica discs were covered with 10 μl of tau extract for 2 min. Afterward, the discs were washed twice with 100 μl double-distilled water (ddH2O) and stored protected from light at RT until imaging. Samples were scanned at a Dimension FastScan with ScanAsyst AFM (Bruker) with a Fastscan-B (Bruker) cantilever in ScanAsyst air mode. Scans were 5 × 5 μm in size, with a scan rate of 3.38 Hz and 1024 samples/line. After acquisition, images were processed with Research NanoScope software (Bruker) by flattening images and adjusting the z-scale from −10 to 20 nm. Tau fibril length was manually analyzed by measuring the distance between two points in Gwyddion (GPL, free software). For each tau extract, 2 to 3 images were analyzed with 50 to 200 fibrils measured per image.
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