Poly-L-lysine (5 µL, 0.01% (w/v), mol wt 7000–150000) was placed on freshly cleaved muscovite mica (Agar Scientific, Stansted, Essex, UK), left for 2 minutes and the mica was then dried in a stream of N2 at room temperature. Viral lysates were diluted (1∶10) in ultrapure water (18 MΩ; Sigma-Aldrich) and placed (5 µL) on the mica surface, left for 2 minutes after which glutaraldehyde (5 µL; 4% (w/v) in 0.2 M sodium cacodylate) was added. After a further 2 minutes, the surface was extensively rinsed with ultrapure water and dried as above. The mica was then mounted on a nickel disc (1 cm diameter) with double-sided adhesive tape and placed on the AFM scanner. AFM studies were performed using a Multi-Mode/NanoScope IV Scanning Probe Microscope (Bruker, Santa Barbara, CA, USA) in air under ambient conditions (T = 24°C, RH = 45%) using the J-scanner (max. xy = 200 µm). Scanning was performed in Tapping Mode using a Si cantilever with integrated tip (tnom = 3.6–5.6 µm, lnom = 140–180 µm, wnom = 48–52 µm, υnom = 200–400 kHz, υmeas = 333.7 kHz, knom = 12–103 Nm−1, Rnom<10 nm; Model: OTESP, Bruker, France) and an RMS amplitude of 2.0 V. Images were subsequently processed (plane fitted and shaded using an artificial light source) using NanoScope analysis software (V 1.40, Bruker, CA, USA).
Multi mode nanoscope 4 scanning probe microscope
Manufactured by Bruker
Sourced in United States
The Multi-Mode/NanoScope IV Scanning Probe Microscope is a high-performance, versatile instrument used for nanoscale imaging and analysis. It combines multiple scanning probe microscopy techniques, including atomic force microscopy (AFM) and scanning tunneling microscopy (STM), to provide detailed surface characterization at the nanometer scale.
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4 protocols using multi mode nanoscope 4 scanning probe microscope
1
Virus Imaging on Mica by AFM
2
Chitosan Nanoparticle Characterization by SEM and AFM
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Chitosan nanoparticles were formulated by ionic gelation [4] . Briefly, OTZ (0.6 mg/mL) was added to chitosan solution (5 mL; 1.75 mg/mL); the solution was stirred for 1 h. TPP solution (2 mL; 1.5 mg/mL) was added drop wise to this solution. This resulted in a particulate dispersion which was placed in an ice bath and sonicated for 30 s. This dispersion was centrifuged at 40,000g for 30 min. The nanoparticles obtained were re-dispersed in water.
Particle size and ζ-potential of the prepared particles were measured at 20 °C using a Malvern Mastersizer 3000HSA, UK. Scanning electron microscopy (SEM) (Zeiss EVO50, Germany) and atomic force microscopy (AFM) (MultiMode/NanoScope IV scanning probe microscope, Santa Barbara, Bruker, USA) were used to characterise the morphology of the prepared particles. AFM experiments were performed, as described elsewhere [5] . The images were then processed and height profiles extracted using NanoScope Analysis software (V 1.4, Bruker, CA, USA). SEMsamples were prepared as follows: One drop of the nanoparticle suspension was placed on a slide cover and left to dry overnight. The slide cover was then attached to an aluminium stud using double-sided adhesive conductive carbon tape. The sample was coated with a layer of gold/ palladium using sputter coating and then analysed under high vacuum at a voltage of 20 kV.
Particle size and ζ-potential of the prepared particles were measured at 20 °C using a Malvern Mastersizer 3000HSA, UK. Scanning electron microscopy (SEM) (Zeiss EVO50, Germany) and atomic force microscopy (AFM) (MultiMode/NanoScope IV scanning probe microscope, Santa Barbara, Bruker, USA) were used to characterise the morphology of the prepared particles. AFM experiments were performed, as described elsewhere [5] . The images were then processed and height profiles extracted using NanoScope Analysis software (V 1.4, Bruker, CA, USA). SEMsamples were prepared as follows: One drop of the nanoparticle suspension was placed on a slide cover and left to dry overnight. The slide cover was then attached to an aluminium stud using double-sided adhesive conductive carbon tape. The sample was coated with a layer of gold/ palladium using sputter coating and then analysed under high vacuum at a voltage of 20 kV.
3
Liposome Characterization by AFM
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Liposome samples were prepared as for SEM, but without Au/Pd coating. Atomic force microscopy (AFM) studies were carried out using a Multi-Mode / NanoScope IV scanning probe microscope, Bruker, Santa Barbara, CA, USA and were performed in air under ambient conditions (T = 23 °C, RH = 21%) using the J-scanner (max. xy = 200 µm). Scanning was performed in Tapping mode using Si cantilevers with integrated tips (t = 3.6-5.6
µm, l = 140-180 µm, w = 48-52 µm, υ0 = 288-338 kHz, k = 12-103 N m -1 , R < 7 nm; model: OTESPA, Bruker, France), and an RMS amplitude of 0.8 V. The images were subsequently processed and dimensions (particle diameters) measured using NanoScope Analysis software (V1.4, Bruker, Santa Barbara, CA, USA).
µm, l = 140-180 µm, w = 48-52 µm, υ0 = 288-338 kHz, k = 12-103 N m -1 , R < 7 nm; model: OTESPA, Bruker, France), and an RMS amplitude of 0.8 V. The images were subsequently processed and dimensions (particle diameters) measured using NanoScope Analysis software (V1.4, Bruker, Santa Barbara, CA, USA).
4
Atomic Force Microscopy of Membrane Entities
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Morphology of unloaded and CP-loaded MEs was observed using AFM. An aliquot of a solution (5 µl) was placed on a surface of freshly-cleaved muscovite mica (1 cm 2 ; Agar Scientific, Stansted, Essex, UK), dried in the desiccator, and further dried in a N 2 stream when needed. The surface was then attached to a nickel disk mounting assembly (1 cm 2 ) using double-sided adhesive tape and placed on top of the AFM scanner. AFM studies were carried out using a Multi-Mode/NanoScope IV scanning probe microscope, Bruker, Santa Barbara, CA, USA and were performed in air under ambient conditions performed in tapping mode using Si cantilevers with integrated tips (t = 3.2-4.2 µm, l = 145-175 µm, w = 38-42 µm, υ 0 = 200-400 kHz, k = 8.4-57 N m -1 , R<10 nm; model: OTESPA-R3, Bruker, France), and an RMS amplitude of 2.0 V. The images were subsequently processed and dimensions measured using NanoScope Analysis software
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