Basement membrane stiffness was measured by Atomic Force Microscopy (AFM)8 (link). Briefly, live follicles were indented four times using a pyramidally-tipped cantilever of calibrated spring constant mounted on a Bruker Catalyst AFM, with an approach velocity of 0.4 μm/sec and a setpoint force of 1nN. Young’s Modulus of elasticity was calculated by fitting the cantilever deflection versus piezo extension curves to the modified Hertz model, using only the first 50 nm of indentation to isolate elasticity from the BM.
Catalyst afm
Manufactured by Bruker
Sourced in United States
The Catalyst AFM is a scanning probe microscope designed for high-resolution imaging and analysis of samples. It utilizes atomic force microscopy (AFM) technology to provide topographical and physical property data on a wide range of materials at the nanoscale. The Catalyst AFM is capable of operating in various imaging modes to accommodate diverse sample types and research needs.
Automatically generated - may contain errors
Lab products found in correlation
Ix51 microscope, by Olympus (1 mentions)
Scanasyst air, by Bruker (1 mentions)
Rtespa, by Bruker (1 mentions)
Axiovert 200, by Zeiss (1 mentions)
Orca r2, by Hamamatsu Photonics (1 mentions)
Matlab, by MathWorks (1 mentions)
Scanasyst fluid, by Bruker (1 mentions)
Dimension icon afm, by Bruker (1 mentions)
Mlct c, by Bruker (1 mentions)
7 protocols using catalyst afm
1
Probing Basement Membrane Elasticity
2
Nanoscale Collagen Fibril Analysis
3
Cellulose Fiber Characterization via AFM
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cellulose fibers were deposited onto freshly cut mica substrates from fiber solutions at 0.1 g L−1 and dry overnight. Fibers were observed by an Olympus IX51 microscope with a 20× objective. Atomic force microscopy imaging was performed with the cantilever located on certain cellulose fibers with the aid of an optical microscope. Topographical images were registered by a Catalyst AFM (Bruker). The images were obtained in tapping mode under ambient air conditions (temperature and relative humidity) using a monolithic silicon tip (Scanasyst-Air, Bruker) with a spring constant of 0.4 N m−1 and a nominal frequency of 70 kHz. Image processing was performed with the WSxM 4.0 software [53 (link)].
4
Evaluating Collagen Fibril Structure
5
Atomic Force Microscopy of Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Measurements were taken on a Catalyst AFM (Bruker Corp.) instrument mounted on the stage of an Axiovert 200 inverted epifluorescence microscope (Zeiss) placed on a vibration isolation table (Isostation). Fluorescence images of the cells were acquired with a cooled CCD camera (Orca R2, Hamamatsu). V-shaped gold-coated silicon nitride cantilevers with four-sided pyramidal tips (MLCT, Bruker Corp.) were used as probes. Nominal spring constant of the cantilevers was , but the actual spring constant of each cantilever was measured before starting the experiments using the thermal fluctuations method.
6
Measuring Effective Elastic Modulus of Cellular Matrices
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The effective elastic modulus of each matrix coated or polymerized in glass-bottom MatTek dishes was measured by performing indentations using a commercial atomic force microscope (AFM) instrument (AFM Catalyst; Bruker). Polystyrene microspheres 5 µm in diameter were attached to tipless cantilevers (MLCT; Bruker) using a UV-curable epoxy adhesive. The spring constants of all cantilevers were estimated using the thermal tune utility of the instrument. For each matrix, indentations were performed over 16 × 16 point grids covering areas of 30 × 30 µm2 that were equal to or larger than a typical cell. A few locations (typically 3–5) lying several millimeters apart from each other were probed. The collected force curves were analyzed using the Hertz model with a custom-written Matlab (MathWorks) software code (supplemental material ). The resulting estimates of effective Young’s modulus for each matrix are presented in Table 1 .
7
Nanomechanical Analysis of sEVs and Cells using AFM
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cantilever of an AFM can
be applied to sense and amplify the
force between the probe on the cantilever beam and the atoms of the
tested samples. Owing to its atomic resolution, we performed a nanomechanical
analysis of living sEVs and GC cells. Dimension Icon AFM (Bruker,
Santa Barbara, CA, USA) can detect sEVs; the type of probe was a ScanAsyst-Fluid
(Bruker, Santa Barbara, CA, USA). In the experiment, we used AFM peak
force QNM mode to image sEVs, which can probe the topography and Young’s
moduli of samples. To detect force curves of cell lines, we utilized
the AFM catalyst (Bruker, Camarillo, CA, USA) and calculated the Young’s
moduli of the cells based on the Hertz model. For experiments related
to TGF-β1 treatment, we employed the AFM JPK NanoWizard (Bruker,
Santa Barbara, CA, USA) to acquire force curves and determined the
Young’s moduli of the cells using the Hertz model. The probe
used for all cell detection was the MLCT-C (Bruker, Santa Barbara,
CA, USA).
be applied to sense and amplify the
force between the probe on the cantilever beam and the atoms of the
tested samples. Owing to its atomic resolution, we performed a nanomechanical
analysis of living sEVs and GC cells. Dimension Icon AFM (Bruker,
Santa Barbara, CA, USA) can detect sEVs; the type of probe was a ScanAsyst-Fluid
(Bruker, Santa Barbara, CA, USA). In the experiment, we used AFM peak
force QNM mode to image sEVs, which can probe the topography and Young’s
moduli of samples. To detect force curves of cell lines, we utilized
the AFM catalyst (Bruker, Camarillo, CA, USA) and calculated the Young’s
moduli of the cells based on the Hertz model. For experiments related
to TGF-β1 treatment, we employed the AFM JPK NanoWizard (Bruker,
Santa Barbara, CA, USA) to acquire force curves and determined the
Young’s moduli of the cells using the Hertz model. The probe
used for all cell detection was the MLCT-C (Bruker, Santa Barbara,
CA, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!