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Mfp 3d bio atomic force microscope

Manufactured by Oxford Instruments
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The MFP-3D Bio Atomic Force Microscope is a high-resolution imaging and analysis instrument designed for biological research. It provides nanoscale topographical and mechanical property measurements of biological samples in liquid environments.

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Lab products found in correlation

Matlab, by MathWorks (4 mentions) Ti u fluorescent inverted microscope, by Nikon (4 mentions) Fluid cell lite coverslip holder, by Oxford Instruments (3 mentions) Axio observer z1, by Zeiss (3 mentions) Ti u fluorescent inverted scope, by Nikon (3 mentions) Mfp 3d bio software, by Oxford Instruments (2 mentions) Pnp tr tips, by NanoWorld (2 mentions) Axiostar plus optical microscope, by Zeiss (1 mentions) Fluo view 1000 confocal laser microscope, by Olympus (1 mentions) Live dead baclight bacterial viability kit, by Thermo Fisher Scientific (1 mentions) Igor pro 6.22a, by Wavemetrics (1 mentions) Igor pro 6.34a, by Oxford Instruments (1 mentions) Ac240ts r3, by Oxford Instruments (1 mentions) Snl probe, by Bruker (1 mentions) Eclipse ti2 microscope, by Nikon (1 mentions) Ac240, by Olympus (1 mentions) Igor pro, by Wavemetrics (1 mentions) Mica surface, by Ted Pella (1 mentions) Ac160ts r3, by Olympus (1 mentions) Vo 200, by Memmert (1 mentions)

23 protocols using mfp 3d bio atomic force microscope

1

Quantifying Bacterial Cell Morphology

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Cells were visualized using an Axiostar plus optical microscope (Carl Zeiss, Germany) in a phase contrast mode. Images were captured with the Pixera PRO 150ES camera (Pixera, USA) and analyzed using VideoTesT-Size 5.0 (Akond, Russia). DCF effects on cell morphology and surface relief were examined by a combined atomic-force and confocal laser scanning system consisting of a MFP-3D-BIO™ atomic force microscope (AFM, Asylum Research Inc., USA), and an Olympus Fluo View 1000 confocal laser microscope (CLSM, Olympus Corporation, Japan). To differentiate living and dead cells, the bacterial suspension was stained with LIVE/DEAD®BacLight™ Bacterial Viability Kit (Invitrogen, USA). The root-mean-square roughness of the cell surface, cell length and width were determined, and the cell volume and surface area were calculated. The images were processed using the Igor Pro 6.22 A (WaveMetrics, USA).
Ivshina I.B., Tyumina E.A., Kuzmina M.V, & Vikhareva E.V. (2019). Features of diclofenac biodegradation by Rhodococcus ruber IEGM 346. Scientific Reports, 9, 9159.
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2

Nanoindentation of Fly Cardiac Muscle

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Adult female flies were anesthetized and immobilized on 25-mm glass coverslips with a thin layer of vacuum grease ventral-side-up. The heart tube was exposed via microsurgery as previously described (Ocorr et al., 2007 (link)) with additional micropipette aspiration to remove all ventral tissue proximal to the conical chamber. Each coverslip is mounted on a Fluid Cell Lite coverslip holder (Asylum Research, Goleta, CA, USA) with 1 mL of hemolymph. Hearts are checked for regular contractions to ensure they are in good health and then resubmerged in 10 mm ethylene glycol tetraacetic acid (EGTA)-treated hemolymph to arrest contraction. Prior to indentation, probes were calibrated via thermal noise method in MFP-3D Bio software (Asylum Research). Nanoindentation was performed on an MFP-3D Bio Atomic Force Microscope (Asylum Research) mounted on a Ti-U fluorescent inverted microscope (Nikon Instruments, Melville, NY, USA) with 120 pN nm−1 silicon nitride cantilevers with premounted, 2-μm radius borosilicate glass spheres (Novascan Technologies, Ames, IA, USA). All indentation curves were analyzed to calculate myocardial stiffness using previously published, automated software custom-written in MATLAB (MathWorks, Natick, MA, USA; Kaushik et al., 2011 (link), 2012 (link)).
Nishimura M., Kumsta C., Kaushik G., Diop S.B., Ding Y., Bisharat-Kernizan J., Catan H., Cammarato A., Ross R.S., Engler A.J., Bodmer R., Hansen M, & Ocorr K. (2014). A dual role for integrin-linked kinase and β1-integrin in modulating cardiac aging. Aging Cell, 13(3), 431-440.
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3

Decellularized hfRPE Substrate Characterization

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After 7 weeks in culture on coverslips, hfRPE cultures were decellularized. An MFP-3D-Bio atomic force microscope (Asylum Research) was utilized to take force maps of 90 × 90 μm. Each of the force maps was made up of a 32 × 32 grid of individual force curves which were acquired using a trigger point of 20 nN and a force distance of 4 μm with a velocity of 4 μm s−1. Force curves were acquired with a polystyrene, 6.1 μm diameter CP-Cont-PS colloidal probe (sQube) and fitted with the Hertz model. Asylum Research's software (Igor Pro 6.34A) was used for the thermal tuning calibrations of the cantilever spring constant.
Benedicto I., Lehmann G.L., Ginsberg M., Nolan D.J., Bareja R., Elemento O., Salfati Z., Alam N.M., Prusky G.T., Llanos P., Rabbany S.Y., Maminishkis A., Miller S.S., Rafii S, & Rodriguez-Boulan E. (2017). Concerted regulation of retinal pigment epithelium basement membrane and barrier function by angiocrine factors. Nature Communications, 8, 15374.
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4

Imaging Biomolecules on Mica Surface

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Samples (50 μl) were applied to a freshly cleaved mica plate. After 1 min, the mica plate was washed gently with water and dried in a stream of argon gas. The samples were imaged in air, at room temperature, under controlled humidity in tapping mode using a MFP-3D-BIO Atomic Force Microscope (Asylum Research, Goleta, CA, USA) equipped with an Olympus AC240TS-R3 (Asylum Research, Goleta, CA) silicon nitride probe (k = ∼1.7 N/m).
Gupta R., Unciuleac M.C., Shuman S, & Glickman M.S. (2016). Homologous recombination mediated by the mycobacterial AdnAB helicase without end resection by the AdnAB nucleases. Nucleic Acids Research, 45(2), 762-774.
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5

Atomic Force Microscopy of Mfp-3S and COL1A1

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An MFP-3D-Bio Atomic Force Microscope (AFM, Asylum Research) was used to obtain images with an SNL probe (Bruker) under tapping mode at room temperature (22 °C). Mfp-3s and COL1A1 was deposited on a mica surface (area ~ 1 cm2) by adsorbing 50 μL of the protein from a 20 μg/ml mfp3s concentration or a 25 μg/ml of COL1A1 concentration in buffer solution at pH 3.0 or 7.5.
Martinez Rodriguez N.R., Das S., Kaufman Y., Wei W., Israelachvili J.N, & Waite J.H. (2015). Mussel adhesive protein provides cohesive matrix for collagen type-1α. Biomaterials, 51, 51-57.
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6

Characterizing Stiffness Gradients in PA Gels

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Stiffness gradient PA gels were mechanically characterized using an MFP-3D-BIO Atomic Force Microscope (Asylum Research), mounted on a Nikon Eclipse Ti2 microscope, in contact mode. Detailed AFM methods are provided in the Supplemental Methods.
Guo T., Jiang C.S., Yang S.Z., Zhu Y., He C., Carter A.B., Antony V.B., Peng H, & Zhou Y. (2023). Mitochondrial fission and bioenergetics mediate human lung fibroblast durotaxis. JCI Insight, 8(1), e157348.
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7

Polyacrylamide Gel Elasticity Mapping

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Atomic Force Microscopy (AFM) measurements of polyacrylamide gels were performed using an MFP-3D-BIO atomic force microscope (Asylum Research, Santa Barbara, CA). Olympus TR400PB AFM probes with an Au/Cr coated silicon nitride cantilever and pyramidal tip were used, with spring constants ranging from 20–30 pN/nm, as measured by thermal calibration. Force maps in square regions of 40 μm edge length, with 169 points per force map, were taken at equal spacing across the gels. Measurements were performed across at least 4 mm length on each side of the interface dividing the primary and secondary ECM regions, as shown in Fig. 1b. Elastic moduli were extracted from force curves using a modified Hertz model [23 (link)].
Nasrollahi S., Walter C., Loza A.J., Schimizzi G.V., Longmore G.D, & Pathak A. (2017). Past matrix stiffness primes epithelial cells and regulates their future collective migration through a mechanical memory. Biomaterials, 146, 146-155.
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8

Stiffness Mapping of Endothelial Cells

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AFM was used to examine the stiffness of HUVECs and adult human adipose ECs. Bright-field images of cells, for determination of the location of stiffness measurements, were acquired using an inverted microscope (Zeiss Axio Observer Z1) as the AFM base (20× 0.8 NA objective). An MFP-3D-BIO Atomic Force Microscope (Asylum Research) was used to collect force maps. A 5-μm borosilicate glass beaded probe (Novascan) with a nominal spring constant of 0.12 N m−1 was used for all measurements. Each force map sampled a 60 μm × 60-μm region, in a 20 × 20 grid of force curves (400 force curves total) under fluid conditions which covered an area of 360 μm2. The trigger point was set to 2 nN with an approach velocity of 5 μm s−1. The force-indentation curves were fit to the Hertz model for spherical tips using the Asylum Research software to determine Young’s modulus, with an assumed Poisson’s ratio value of 0.45 for the sample. Force maps of stiffness along with individual stiffness values for each measured point were then exported from the Asylum Research software for further analysis. A custom-made MATLAB (MathWorks) script was written to correctly analyse the data for the stiffness of the cells and filter measurements such that only data 1 μm from the glass bottom dish were analysed (to remove any substrate effect from the measurements).
Palikuqi B., Nguyen D.H., Li G., Schreiner R., Pellegata A.F., Liu Y., Redmond D., Geng F., Lin Y., Gómez-Salinero J.M., Yokoyama M., Zumbo P., Zhang T., Kunar B., Witherspoon M., Han T., Tedeschi A.M., Scottoni F., Lipkin S.M., Dow L., Elemento O., Xiang J.Z., Shido K., Spence J.R., Zhou Q.J., Schwartz R.E., De Coppi P., Rabbany S.Y, & Rafii S. (2020). Adaptable haemodynamic endothelial cells for organogenesis and tumorigenesis. Nature, 585(7825), 426-432.
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9

Membrane Topography Analyzed by AFM

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The cells after MHIFU irradiation with TSL in the PDMS cavity were treated by glutaraldehyde (2.5%, 10min), washed by PBS and then air-dried. AFM imaging was performed on a MFP-3D bio-atomic force microscope (Asylum Research, Santa Barbara, CA) operated in a contact mode. The topographic images of the membrane were acquired at a scan field of 5 μm×5 μm.
Meng L., Deng Z., Niu L., Li F., Yan F., Wu J., Cai F, & Zheng H. (2015). A Disposable Microfluidic Device for Controlled Drug Release from Thermal-Sensitive Liposomes by High Intensity Focused Ultrasound. Theranostics, 5(11), 1203-1213.
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10

Probing Myocardial Stiffness in Adult Flies

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All indentation was performed on MFP-3D Bio Atomic Force Microscope (Asylum Research) mounted in a Ti-U fluorescent inverted scope (Nikon Instruments, Melville, NY) with 2 µm radius borosilicate glass beads mounted on a 120 pN/nm silicon nitride cantilevers (Novascan Technologies). All probes were calibrated for precise spring constant via the thermal noise method. Adult female flies with exposed beating hearts were immobilized on 25 mm glass cover slip, which is mounted on a Fluid Cell Lite coverslip holder (Asylum Research) with 1 mL of hemolymph. Hearts were checked for regular contractions and arrested with 10 mM EGTA just prior to indentation. 1 µm2 area was probed with a 4 × 4 grid of indentations and an indentation velocity of 1 µm/s resulting in 16 force curves per replicate. All indentation curves were analyzed to by an automated software custom written in MATLAB (Mathworks) as previously described in [13 (link), 14 (link)] to calculate the elastic modulus (Pa, Pascal), or stiffness, of the myocardium. This software was further adapted to analyze the elastic modulus (Pa) of the ECM between the VM and myocardium and the thickness of this layer in µm. This analysis method can detect the presence of a third material not fit by either the ventral or cardiac fits. A third region is found if R2 > 0.95 and stiffness is 25% less than the VM layer.
Sessions A.O., Kaushik G., Parker S., Raedschelders K., Bodmer R., Van Eyk J.E, & Engler A.J. (2016). Extracellular Matrix Downregulation in the Drosophila Heart Preserves Contractile Function and Improves Lifespan. Matrix biology : journal of the International Society for Matrix Biology, 62, 15-27.
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