Nanowizard 4a bioscience afm

Manufactured by Bruker

The NanoWizard 4a Bioscience AFM is an atomic force microscope that allows for high-resolution imaging and analysis of biological samples. Its core function is to provide detailed topographical information and nanoscale resolution of surface features.

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Lab products found in correlation

Polystyrene microbeads, by Polysciences (1 mentions) Saa sph 5um probe, by Bruker (1 mentions) Data processing software, by Bruker (1 mentions) Jpk data processing software, by Bruker (1 mentions) Lsm5 confocal fluorescence microscope, by Zeiss (1 mentions)

Close spelling variants of this product

NanoWizard 4a AFM Nanowizard 4a NanoWizard AFM

3 protocols using nanowizard 4a bioscience afm

Colloidal Tip Cantilever Force Spectroscopy

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Imaging and force spectroscopy were all performed in liquid with NanoWizard 4a Bioscience AFM (JPK Instruments). Imaging was done using qp-BioAC-Cl-10 cantilevers (Nanosensors) and CB2 tip with a measured stiffness of 0.099 N m^–1. Force spectroscopy measurements were done with custom colloidal tip cantilevers at room temperature and 50 °C with 15 minutes of settling time. Polystyrene microbeads (6 μm, Polysciences) were dispersed onto a glass slide, dried and bound to cantilevers (HQ:CSC37, MikroMasch) using a glue (M-Bond 610 adhesive Kit, SPI). The colloidal tip cantilever had a measured stiffness of 0.374 N m^–1. Force curves were obtained on 16 microactuators and each indentation was performed three times. Prior to both imaging and force spectroscopy, samples were transferred to a sugar solution and pipetted on to poly-lysine pretreated plastic Petri dishes to facilitate immobilization onto the surface. Each indentation measurement was analyzed by JPK data processing software using the standard Young's modulus calculation approach offered.

Özkale B., Parreira R., Bekdemir A., Pancaldi L., Özelçi E., Amadio C., Kaynak M., Stellacci F., Mooney D.J, & Sakar M.S. (2019). Modular soft robotic microdevices for dexterous biomanipulation †Electronic supplementary information (ESI) available. See DOI: 10.1039/c8lc01200h. Lab on a Chip, 19(5), 778-788.

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Characterizing Gelatin Microcarrier Mechanics

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The Young’s modulus and topology of the gelatin microcarriers were measured in 1 × PBS with 0.1% Tween 80 using a JPK Nanowizard 4a BioScience AFM in force spectroscopy mode. The microcarriers were adhered to a poly-lysine coated slide (Epredia) and indented with a SAA-SPH-5UM probe (Bruker) with a 10 μm diameter spherical tip. The spring constants of the probes were individually calibrated by the manufacturer. Single indentations were performed with a total force of 4.0 nN. Since an oblique contact between the spherical AFM probe and the microcarrier surface can result in inaccurate force curve fitting, indentations were performed on the top surface of the microcarriers (the top of the ridges of the gMCs or the apex of the sMCs). Young’s modulus values were determined by averaging over 5 unique indentations for sMCs and over at least 15 unique indentations along the top of multiple parallel ridges for gMCs. All force curve analysis was performed using the JPK Data Processing software. The Young’s modulus was calculated by using a Hertz/Sneddon spherical fit with a Poisson’s ratio of ν = 0.5 [43 (link)].

Norris S.C., Kawecki N.S., Davis A.R., Chen K.K, & Rowat A.C. (2022). Emulsion-templated microparticles with tunable stiffness and topology: Applications as edible microcarriers for cultured meat. Biomaterials, 287, 121669.

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Quantifying Nuclear Stiffness by AFM

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SMCs in 35 mm glass coverslip-bottom dishes (World Precision Instruments) were incubated with Dox for 24 hours to induce nuclear lamin expression. Nuclear stiffness (Young’s modulus) in adherent cells was measured on a NanoWizard 4a Bioscience AFM (JPK Instruments) coupled with a Zeiss LSM5 confocal fluorescence microscope (53 (link), 54 (link)). A spherical AFM tip with a radius of 500 nm and 0.2 N/m spring constant cantilever (Nanotools, B500-CONT) was placed on top of the cell over the nucleus, identified by staining DNA with Hoechst 33342. Three different areas of the nucleus were sampled with a 2-nN maximum set point. The 3 measurements were used to calculate the average Young’s modulus for a single cell. All measurements were performed at 37° C in HEPES-buffered culture medium. The Young’s modulus was measured in 25 randomly selected cells for each group, and the force curves analyzed with JPK Data Processing software (JPK Instruments). The Young’s modulus was calculated with the Hertz model for a spherical tip and applied to fit the slopes of the approach curve.

Kim P.H., Chen N.Y., Heizer P.J., Tu Y., Weston T.A., Fong J.L., Gill N.K., Rowat A.C., Young S.G, & Fong L.G. (2021). Nuclear membrane ruptures underlie the vascular pathology in a mouse model of Hutchinson-Gilford progeria syndrome. JCI Insight, 6(16), e151515.

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