Sample preparation for AFM imaging was performed as previously described[16 (link)]. Freshly cleaved mica was functionalized with a solution of 1-(3-aminopropyl)- silatrane (APS) for sample deposition[23 (link)]. The samples were diluted from 300 nM to 2 nM in imaging buffer (10 mM HEPES pH 7.5, 4 mM MgCl2) immediately before deposition on the functionalized mica. The sample was left to incubate for 2 minutes before being rinsed with water and dried with argon flow. Samples were stored in vacuum and argon before being imaged on Multimode AFM/Nanoscope IIId system using TESPA probes (Bruker Nano Inc, Camarillo, CA). A typical image captured was 1×1 μm in size with 512 pixels/line.
Multimode afm nanoscope iiid system
Manufactured by Bruker
Sourced in United States
The Multimode AFM/Nanoscope IIId system is a high-performance atomic force microscope (AFM) designed for advanced materials characterization. The system provides nanoscale imaging, measurement, and manipulation capabilities, allowing users to investigate the surface topography, mechanical, and electrical properties of a wide range of samples.
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5 protocols using multimode afm nanoscope iiid system
1
Sample Preparation for AFM Imaging
2
Nucleosome Sample Preparation for AFM Imaging
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Sample preparation for AFM imaging was performed as previously described26 (link). Freshly cleaved mica was functionalized with a solution of 1-(3-aminopropyl)- silatrane (APS). The nucleosome stock solution was diluted from 300 to 2 nM in imaging buffer (10 mM HEPES pH 7.5, 4 mM MgCl2) immediately before deposition on the functionalized mica. The sample was left to incubate for 2 min before being rinsed with water and dried with argon flow. Samples were stored in vacuum before being imaged on Multimode AFM/Nanoscope IIId system using TESPA probes (Bruker Nano Inc, Camarillo, CA). A typical image captured was 1 × 1 μm in size with 512 pixels/line.
3
Nucleosome Deposition on APS-Modified Mica
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A 167 μM solution of 1-(3-aminopropyl)- silatrane (APS) was used to modify freshly cleaved mica for 30 minutes at RT as previously described (23 (link),24 (link)). The nucleosome stock solution was diluted to 2 nM (based on DNA concentration) in a buffer containing 10 mM HEPES (pH 7.5) and 4 mM MgCl2. Immediately following nucleosome dilution, 7 μl of sample was deposited on chilled (4°C) APS-mica and was rinsed with 1.5 ml of ultrapure H2O after a 2-min incubation which was then followed by drying with a light flow of argon air. Prepared samples were stored under vacuum until being imaged at ambient temperature and humidity conditions on a Multimode AFM/Nanoscope IIId system using TESPA probes (Bruker Nano Inc). A typical image captured was 1.5 × 1.5 μm in size with 512 pixels/line.
4
Sample Preparation for Nucleosome AFM
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Sample preparation for AFM imaging was performed as previously described [24 (link)]. Freshly cleaved mica was functionalized with a solution of 1-(3-aminopropyl)- silatrane (APS). The nucleosome stock solution was diluted from 300 nM to 2 nM in imaging buffer (10 mM HEPES pH 7.5, 4 mM MgCl2) immediately before deposition on the functionalized mica. The sample was left to incubate for 2 min before being rinsed with water and dried with argon flow. Samples were stored in vacuum before being imaged on a Multimode AFM/Nanoscope IIId system using TESPA probes (Bruker Nano Inc., Camarillo, CA, USA). A typical image captured was 1 × 1 μm in size with 512 pixels/line.
5
Atomic Force Microscopy Characterization of Nanoparticles
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Samples for AFM scanning were diluted for each conjugation method to 0.5 nM before depositing onto 1-(3-aminopropyl)-silatrane (APS) functionalized mica. The samples were incubated for 2 min to bind to the APS mica, followed by gentle washing with DI water and drying with a slow argon gas flow. The samples were then dried in a vacuum chamber overnight.
Once dried, the samples were scanned on a Multimode AFM/Nanoscope IIId system with TESPA probes (Bruker Nano Inc., Camarillo, CA). The images captured were 3 × 3 μm in size with 1536 pixels/line.
The images were analyzed using Femtoscan software (Advance Technologies Center, Moscow, Russia). A cross-sectional line was drawn over the center of each particle and a diameter in nm was obtained. Once all the particles on each image were analyzed, histograms were generated using Origin software.
Once dried, the samples were scanned on a Multimode AFM/Nanoscope IIId system with TESPA probes (Bruker Nano Inc., Camarillo, CA). The images captured were 3 × 3 μm in size with 1536 pixels/line.
The images were analyzed using Femtoscan software (Advance Technologies Center, Moscow, Russia). A cross-sectional line was drawn over the center of each particle and a diameter in nm was obtained. Once all the particles on each image were analyzed, histograms were generated using Origin software.
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