FastScan-D probes (Bruker), nominal spring constant 0.25 N m-1, resonant frequency (in liquid) 110 kHz were used for all presented images taken in liquid. Tespa-V2 probes (Bruker), nominal spring constant of 37 N m-1, resonant frequency (in air) 320 kHz, were used for all experiments in air.
Fastscan d probes
Manufactured by Bruker
Sourced in United States
FastScan-D probes are high-performance atomic force microscopy (AFM) probes designed for fast and high-resolution imaging. They feature a proprietary design that enables rapid scanning and high-quality data acquisition.
Automatically generated - may contain errors
Lab products found in correlation
Tespa v2 probes, by Bruker (2 mentions)
Nanoscope iiia controller, by Digital Instruments (2 mentions)
Fast scan bio afm, by Bruker (2 mentions)
Multimode afm, by Digital Instruments (2 mentions)
Tr400psa silicon nitride probes, by Olympus (2 mentions)
Dimension fastscan afm, by Bruker (1 mentions)
Multimode afm, by Bruker (1 mentions)
Cypher es afm, by Oxford Instruments (1 mentions)
Fastscan bio, by Bruker (1 mentions)
11 protocols using fastscan d probes
1
Atomic Force Microscopy Probe Specifications
2
Atomic Force Microscopy of Polymer Envelopes
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Imaging was carried out using a Bruker Dimension FastScan AFM using FastScan-D probes (Bruker—nominal k = 0.25 N/m, nominal cantilever length = 16 μm) in “Tapping Mode” (Amplitude Modulated Intermittent Contact Mode) driven at ~110 kHz with a free amplitude of ~1 nm.
All imaging of polymer envelopes (sacculi) was carried out in liquid (buffer) without drying at any point.
All imaging of polymer envelopes (sacculi) was carried out in liquid (buffer) without drying at any point.
3
Imaging DNA Hybrids via AFM
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AFM imaging was performed in photothermal off-resonance tapping45 (link) on a custom-built microscope based on a Bruker MultiMode AFM, as describe elsewhere46 . DNA hybrids were diluted in 10 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) pH 7.3 to a concentration of about 5 ng/µL. Two microliters of 20 mM NiCl2 solution was pipetted on a freshly cleaved 3 mm Mica disk. Then, 2 µL of 10 mM HEPES pH 7.3 solution was added. Finally, 500 nL of the diluted DNA solution was added. The sample was left to incubate for several minutes to let the DNA adhere to the Mica. Afterwards the scan head with a hanging droplet of 10 mM HEPES supplemented with 2 mM NiCl2 was added on top of the scanner for imaging. Images were recorded using Bruker FastScan-D probes at 30 kHz PORT rate and at 10 lines/s.
4
Atomic Force Microscopy Imaging of Biomolecules
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1-5 μl of transcription product was mixed with 40 μl AFM dilution buffer (12.5 mM Mg(OAc)2, 40mM KCl, 40mM NaCl, Tris-Borate pH 7.8) directly on the surface of a freshly-cleaved mica puck. Mixing is performed by vigorously pumping a 200 μl pipette tip ten times, before removing and discarding the fluid. The mica was washed with a solution of 60 mM NiCl2. Most AFM images were collected using a Multimode AFM (Digital Instruments) with a Nanoscope IIIA controller and a J-scanner. Olympus TR400PSA silicon nitride probes with a spring constant of ~.08 N/m were used for imaging, with a drive frequency of ~6-9 kHz. AFM in Supplementary Figs. 10 and 11 were collected with a Bruker Fastscan Bio AFM (Bruker) under buffer using FastScan-D probes (Bruker).
5
High-Resolution Imaging of Bacterial Sacculi
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Unless otherwise stated data were collected on this instrument. Live cell imaging was carried out in “Tapping Mode” under a buffer comprised of 300 mM KCl, 10 mM Tris, pH 7.8 using Bruker FastScanD probes driven at ~80 kHz with a typical free amplitude of ~1-2 nm and a set-point amplitude typically ~60-90% of the free amplitude35 (link). This corresponds to average tip-sample forces ~ 100 pN. Imaging of the external surface of sacculi at high resolution with “Tapping Mode” was not achieved, possibly because of the diffuse nature of the surface when relaxed. Sacculi imaging was instead carried out in “PeakForce Tapping” mode36 with a typical amplitude of 100-200 nm, frequency of 2-8 kHz and setpoint force of 1-2 nN with buffer conditions which varied across the samples between HPLC water and 150-300 mM KCl +10mM Tris (pH 8). All images are topographic images unless otherwise stated. All 3D visuals were created using Nanoscope Analysis software.
6
Atomic Force Microscopy Probe Specifications
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FastScan-D probes (Bruker), nominal spring constant 0.25 N m-1, resonant frequency (in liquid) 110 kHz were used for all presented images taken in liquid. Tespa-V2 probes (Bruker), nominal spring constant of 37 N m-1, resonant frequency (in air) 320 kHz, were used for all experiments in air.
7
High-Resolution Imaging of G-Wires via Atomic Force Microscopy
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Experiments were performed using a Cypher ES AFM purchased from Asylum Research (Oxford Instruments). Solution AFM images were obtained in amplitude modulated ac mode (commonly known as tapping mode) using FastScan-D probes (Bruker) having a cantilever of 16 µm length and 4 µm width. Biolever mini AFM probes (from Asylum Research) were used for a few initial trials but did not allow reproducible high-resolution imaging of G-wires. Experiments were performed at 10 °C (via sample stage cooling), at which the FastScan-D probes had a resonant frequency of 90–100 kHz, which was slightly lower than the frequency at room temperature. A pulsed blue laser (BlueDrive) with ~ 5 µm circular spot size was focused at the base of the cantilever for photothermal excitation51 . For detecting cantilever deflections, a superluminescent diode (SLD) laser with 3 × 9 µm spot size was used. Cantilever amplitudes of < 1 nm with imaging speeds of 10,000 pixels/s were used (only) for high-resolution imaging.
8
AFM Imaging of DNA Nanostructures
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After being diluted to 1 nM (scaffold concentration) in 1 × TE/Mg2+ buffer, 40 μL of each sample was deposited onto freshly cleaved mica (SPI Supplies, catalog # 01873-CA). The solution was removed after 30 s, the mica surface was washed three times with 40 μL TE buffer containing 10 mM MgCl2 and 100 mM NaCl, and 80 μL of 1 × TE/Mg2+ buffer was then added before imaging. Imaging was done in fluid using FastScan-D probes (Bruker) and tapping mode on a FastScan Bio (Bruker), typically at a scan rate of 5 Hz with 1024 lines per image. The amplitude setpoint was usually between 30 and 50 mV, with drive amplitude at 180 to 240 mV and drive frequency at 110 Hz. The integral and proportional gains were set to 1 and 2, respectively. Samples that were not imaged immediately after the fluorescence kinetics experiments were kept at −20 °C and thawed before AFM imaging.
9
High-Resolution Imaging of Bacterial Sacculi
10
Functionalized Mica Imaging using AFM
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Functionalized mica was prepared, and samples were deposited as described above. A measure of 10 mM Tris-HCl pH 8.0 buffer was used for liquid imaging. The AFM used in this work is a Nanoscope V system modified to incorporate a small cantilever head during an open-science workshop at EPFL in the laboratory of Professor Georg Fantner [21 (link)]. Samples were imaged using photothermal off-resonance tapping (PORT) [22 (link)] using FASTSCAN-D probes (Bruker, Santa Barbara, CA, USA). Captured images were 500 × 500 nm in size at a resolution of 1 nm/pixel.
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