The fabricated sample surfaces were characterized by AFM in air using an Agilent 5500 and a JPK Nanowizard 3 AFM operated in intermittent contact mode and HQ:NSC18/AlBS cantilevers (MikroMasch, Wetzlar, Germany) with a nominal tip radius <8 nm. In order to minimize the impact of artefacts resulting from tip-to-tip variations and tip wear, cantilevers were frequently replaced by fresh ones during the course of the AFM measurements, so that the results of the statistical analyses represent not only averages of several AFM images but also of several cantilevers. All images were recorded with a scan size and a resolution of 2 × 2 µm² and 1024 × 1024 pixels, respectively, and analyzed using Gwyddion open source software [37 (link)]. To this end, the images were preprocessed by mean plane subtraction, row alignment using the median, and subtraction of a third-degree polynomial. The height values of the images were subsequently normalized by setting the height minimum to zero. Statistical analyses were then carried out using the Statistical Quantities, the Statistical Functions, and the Fractal Dimension tools of Gwyddion. See the Gwyddion user guide for details [38 ].
Hq nsc18 al bs cantilevers
Manufactured by MikroMasch
Sourced in Germany
HQ:NSC18/Al BS cantilevers are silicon-based atomic force microscopy (AFM) probes designed for high-quality imaging. They feature an aluminum backside coating and are suitable for a variety of AFM techniques.
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6 protocols using hq nsc18 al bs cantilevers
1
AFM Characterization of Fabricated Surfaces
2
Atomic Force Microscopy Imaging Protocol
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AFM Imaging was done as described previously13 (link) except for the AFM images of Fig. 2b (wt Redβ, C3Redβ, N1Redβ), which were recorded dry using an Agilent 5100 AFM and HQ:NSC18/Al BS cantilevers (MikroMasch) in intermittent contact mode. All data analysis and data fitting was done in IGOR Pro version 5.04B (WaveMetrics Inc.). The refined models (Fig. 2D ) were generated employing computer-aided design (CAD) construction software SolidWorks 2005 SPO.0 (SolidWorks Corporation) and by the support of Quindium GmbH & Co. KG.
3
Atomic Force Microscopy Imaging Protocol
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The dry samples were imaged in air using a Bruker Dimension ICON (Bruker France S.A.S., Wissembourg, France) in ScanAsyst Peak-Force Tapping mode with ScanAsyst-Air cantilevers (Bruker AFM Probes, Camarillo, CA, USA) and a JPK Nanowizard III (JPK Instruments, Berlin, Germany) in intermittent contact mode with HQ:NSC18/Al BS cantilevers (MikroMasch, Wetzlar, Germany). The obtained AFM images were flattened and height-adjusted using Gwyddion 2.52 open-source software [49 (link)].
4
Atomic Force Microscopy Surface Roughness Measurement
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AFM imaging was performed using a JPK Nanowizard III (Bruker GmbH, Billerica, MA, USA) equipped with an acoustic enclosure in alternating contact mode operating in ambient air conditions at a scan rate of 0.5–1.2 Hz. HQ:NSC18/AlBS cantilevers (75 kHz and 2.8 Nm−1, nominal radius of 8 nm, MikroMasch, Tallinn, Estonia) were used. For subsequent data processing and calculation of RMS roughness values for estimation of surface roughness, the Gwyddion open-source software [48 (link)] (V 2.56, Gwyddion) was employed. RMS roughness measurements were obtained from the average of the roughness values collected from three different 5.0 × 5.0 µm2 areas present in three equally prepared samples.
5
DNA Origami Imaging Using AFM
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For AFM imaging, a 5 μL droplet of
each DNA origami sample (10 nM in 1× TAE with 10 mM MgCl2) with or without 20 μM MB was deposited on a freshly
cleaved mica substrate. After adding 100 μL of 1× TAE with
10 mM MgCl2 to spread the sample over the whole substrate
surface, the sample was incubated for 5–15 min, subsequently
dipped in HPLC-grade water, and dried in a stream of ultrapure air.
AFM imaging was performed in air using Agilent 5100 AFM in intermittent
contact mode and HQ:NSC18/Al BS cantilevers from MikroMasch.
each DNA origami sample (10 nM in 1× TAE with 10 mM MgCl2) with or without 20 μM MB was deposited on a freshly
cleaved mica substrate. After adding 100 μL of 1× TAE with
10 mM MgCl2 to spread the sample over the whole substrate
surface, the sample was incubated for 5–15 min, subsequently
dipped in HPLC-grade water, and dried in a stream of ultrapure air.
AFM imaging was performed in air using Agilent 5100 AFM in intermittent
contact mode and HQ:NSC18/Al BS cantilevers from MikroMasch.
6
Atomic Force Microscopy of Protein-Heparin Complexes
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For atomic force microscopy (AFM) imaging,
a 5 μL droplet of PCS10 or protein copolymer–heparin
complexes in PB or PBS was deposited on a freshly cleaved mica substrate.
Heparin in both media was 0.1 mg mL–1. Protein copolymers
were used to fully neutralize heparin (phosphate buffer (PB)/PCS0
= 4 mg mL–1, PCS4 = 5 mg mL–1,
PCS10 = 7 mg mL–1, and PS = 0.2 mg mL–1; phosphate buffered saline (PBS)/PCS0 = PCS4 = PCS10 = 10 mg mL–1 and PS = 0.3 mg mL–1). PCS10 with
the same concentration in the absence of heparin was also imaged accordingly.
The sample was incubated on mica for 5–15 min, subsequently
dipped in HPLC-grade water, and dried in a stream of ultrapure air.
AFM imaging was performed in the air using Agilent 5100 AFM in intermittent
contact mode and HQ:NSC18/Al BS cantilevers from MikroMasch.
a 5 μL droplet of PCS10 or protein copolymer–heparin
complexes in PB or PBS was deposited on a freshly cleaved mica substrate.
Heparin in both media was 0.1 mg mL–1. Protein copolymers
were used to fully neutralize heparin (phosphate buffer (PB)/PCS0
= 4 mg mL–1, PCS4 = 5 mg mL–1,
PCS10 = 7 mg mL–1, and PS = 0.2 mg mL–1; phosphate buffered saline (PBS)/PCS0 = PCS4 = PCS10 = 10 mg mL–1 and PS = 0.3 mg mL–1). PCS10 with
the same concentration in the absence of heparin was also imaged accordingly.
The sample was incubated on mica for 5–15 min, subsequently
dipped in HPLC-grade water, and dried in a stream of ultrapure air.
AFM imaging was performed in the air using Agilent 5100 AFM in intermittent
contact mode and HQ:NSC18/Al BS cantilevers from MikroMasch.
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