Fibrin clots prepared from plasma of 10 randomly-selected patients as described above were analyzed by SEM. Briefly, clots were fixed using 2.5% glutaraldehyde, washed with distilled water, dehydrated in graded water-ethanol solutions, dried by the critical point procedure, and sputter-coated with gold (14 (link)). Samples were scanned in 10 different areas (microscope JEOL JCM-6000; JEOL Ltd., Tokyo, Japan) at 5,000× and 10,000× magnification to determine a fibrin fiber diameter of at least 50 individual fibers per clot (n=50) within the fibrin network using ImageJ software (US National Institutes of Health, Bethesda, MD, USA). SEM images were analyzed manually by two independent investigators who were unaware of the origin of the samples.
Jcm 6000
Manufactured by JEOL
Sourced in Japan, United States
The JCM-6000 is a compact scanning electron microscope (SEM) designed for high-resolution imaging of a wide range of samples. It features a high-performance electron optical system, sophisticated control software, and a user-friendly interface. The JCM-6000 is capable of producing high-quality images with a resolution of up to 3 nanometers.
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Lab products found in correlation
Smart coater, by JEOL (6 mentions)
Iraffinity 1, by Shimadzu (4 mentions)
Jed 2300, by JEOL (3 mentions)
Sc7620, by Quorum Technologies (3 mentions)
Jfc 1200 fine coater, by JEOL (3 mentions)
Ez lx universal tester, by Shimadzu (2 mentions)
Sc7620 mini sputter coater, by Quorum Technologies (2 mentions)
Dii 29010sctr smart coater, by JEOL (2 mentions)
Jfc 1200, by JEOL (1 mentions)
Human thrombin, by Merck Group (1 mentions)
Tristar 2 3020, by Micromeritics (1 mentions)
Jem 2010, by JEOL (1 mentions)
Tm 1000, by Hitachi (1 mentions)
Msp 1s, by Hitachi (1 mentions)
Smartlab, by Rigaku (1 mentions)
Quorum sc7620 mini sputter coater, by Quorum Technologies (1 mentions)
Quorum k850, by Quorum Technologies (1 mentions)
Cm1950, by Leica (1 mentions)
Quorum sc7620, by Quorum Technologies (1 mentions)
Q150 res, by JEOL (1 mentions)
98 protocols using jcm 6000
1
Fibrin Clot Structural Analysis
2
Characterization of Fibrin Clot Ultrastructure
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After the permeability measurement, fibrin clots were fixated in 2.5% glutaraldehyde (in 0.1 M phosphate-buffered saline solution, pH 7.4) for 2 h. Fixed clots were gently removed from tubes, dehydrated in graded ethanol solutions, dried by the critical point procedure, and sputter coated with gold. Finally, the treated clots were scanned in ten different areas (microscope JEOL JCM6000; JEOL Ltd., Tokyo, Japan). We analyzed fibrin clots from 20 randomly selected AS patients with similar fibrinogen levels, including 10 with the highest protein carbonyl (PC) content (4th quartile, >3.05 nmol/mg) and 10 with lower PC levels (1st–3rd quartiles). For each clot, 4 micrographs were performed at a machine magnification of 10,000 × in order to evaluate fibrin diameter based on 40–50 fibers per clot, using ImageJ software (US National Institutes of Health, Bethesda, MD, USA).
3
Scanning Electron Microscopy of Plasma Fibrin Clots
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Plasma fibrin clots from randomly selected patients and controls were analyzed. Fixation was performed after the permeability measurement with the use of 2.5% of glutaraldehyde in phosphate-buffered saline solution for 2 hours. Fixed clots were gently removed from tubes, washed with distilled water, and then dehydrated in graded water-ethanol solutions, dried by the critical point procedure, and sputter coated with gold. Samples were scanned in six different areas (microscope JEOL JCM-6000; JEOL Ltd., Tokyo, Japan).
4
SEM Imaging of UBM Particulate
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A thin layer of UBM particulate was mounted on adhesive metallic tape and sputter coated with 3.5 nm of gold. The samples were viewed with a JEOL JCM-6000 (JEOL Ltd.) SEM at magnifications ranging from 20× to 500×.
5
Scanning Electron Microscopy of Samples
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Microscope observations were performed in accordance with the method previously reported by Rosicka-Kaczmarek et al. [61 (link)]. Microscopic images were recorded using a Jeol-JCM-6000 scanning electron microscope (Akishima, Japan). The examined samples were sputter-coated with gold under vacuum (without any noble gas), and images were recorded at various acceleration potentials ranging from 5 kV to 10 kV.
6
Fiber Morphology Analysis via SEM
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A benchtop scanning
electron microscope (JEOL JCM-6000, JEOL Ltd., Tokyo, Japan) was used
to investigate the fiber morphology. Fibers were fixed to the sample
holder using conductive carbon stickers (Agar Scientific Ltd., Essex,
UK). The samples were gold-coated in an argon atmosphere using a fine
coater (JEOL JFC-1200, JEOL Ltd.). SEM micrographs were collected
at various magnifications from numerous sites in each sample.
electron microscope (JEOL JCM-6000, JEOL Ltd., Tokyo, Japan) was used
to investigate the fiber morphology. Fibers were fixed to the sample
holder using conductive carbon stickers (Agar Scientific Ltd., Essex,
UK). The samples were gold-coated in an argon atmosphere using a fine
coater (JEOL JFC-1200, JEOL Ltd.). SEM micrographs were collected
at various magnifications from numerous sites in each sample.
7
Plasma Fibrin Clot Structural Analysis
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Plasma fibrin clots from five randomly selected patients classified to SAVR or TAVI were analyzed. Fixation was performed after the permeability measurement using 2.5% glutaraldehyde in phosphate-buffered saline solution for 2 hours. Fixed clots were, washed with distilled water, and then dehydrated in graded ethanol solutions, dried by the critical point procedure, and sputter coated with gold. Samples were scanned in six different areas (microscope JEOL JCM-6000; JEOL Ltd., Tokyo, Japan). Images were analyzed in ImageJ (US National Institutes of Health, Bethesda, MD, USA).
8
Fibrin Clot Permeability and Morphology
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Permeation of plasma fibrin clots was determined as described [27] . In brief, 20 mM calcium chloride and 1 U/ mL human thrombin (Sigma-Aldrich, St. Louis, MO, USA) were added to citrated plasma. Tubes containing the clots were connected to a reservoir of a Tris-buffered saline, and its volume flowing through the gels was measured. A permeation coefficient (K s ), which indicates the pore size, was calculated from the equation:
where Q is the flow rate in time t, L is the length of a fibrin gel, η is the viscosity of liquid, t is percolating time, A is the cross-sectional area, and Δp is a differential pressure.
After K s measurement, clots (n = 6) were fixed using 2.5% glutaraldehyde, then removed from tubes, washed with distilled water, dehydrated in graded water-ethanol solutions, dried by the critical point procedure, and sputter coated with gold. Samples were scanned in six different areas (microscope JEOL JCM-6000; JEOL Ltd., Tokyo, Japan) at magnification of 5,000x to determine a fibrin diameter of at least 50 fibers per clot using the ImageJ software (US National Institutes of Health, Bethesda, MD, USA).
where Q is the flow rate in time t, L is the length of a fibrin gel, η is the viscosity of liquid, t is percolating time, A is the cross-sectional area, and Δp is a differential pressure.
After K s measurement, clots (n = 6) were fixed using 2.5% glutaraldehyde, then removed from tubes, washed with distilled water, dehydrated in graded water-ethanol solutions, dried by the critical point procedure, and sputter coated with gold. Samples were scanned in six different areas (microscope JEOL JCM-6000; JEOL Ltd., Tokyo, Japan) at magnification of 5,000x to determine a fibrin diameter of at least 50 fibers per clot using the ImageJ software (US National Institutes of Health, Bethesda, MD, USA).
9
Characterization of Dragline Silk Morphology and Tensile Properties
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The surface morphology of the dragline silks was observed by SEM (JCM 6000, JEOL Ltd, Tokyo Japan). Samples were mounted on an aluminium stub with conductive tape backing and sputter-coated with gold for 1 min using a Smart Coater (JEOL) prior to SEM visualization at 5 kV. At least eight individual mechanical stretching tests were performed for each dragline silk. The experimental set-up was similar to those reported previously [28 (link)]. Each fibre was attached to a rectangular piece of cardboard with a 5 mm aperture using 95% cyanoacrylate. The tensile properties of the fibres were measured using an EZ-LX universal tester (Shimadzu, Kyoto, Japan) with a 1 N load cell at a strain rate of 10 mm min−1 (0.033 s−1) at 25°C and 48% relative humidity. For each tensile test, the cross-sectional area of an adjacent section of the fibre was calculated based on the SEM images.
10
Characterizing HA-PLLA Membrane Structure
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A wet HA-PLLA composite membrane was fractured after freezing in liquid nitrogen. The membrane was coated with carbon using a carbon coater (CC-50, Shimadzu). The cross-section of the membrane was observed by scanning electron microscopy with energy dispersive X-ray spectroscopy (EDS; JCM-6000 with JED-2300, JEOL).
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