Three live specimens of D. latus were cut into suitably small pieces, fixed in cold (4 °C) 1.5% glutaraldehyde and 1.5% paraformaldehyde solutions in 0.1 M Hepes (pH 7.4) and stored at 4 °C. After washing with 0.1 M Hepes (pH 7.4), they were post-fixed in cold (4 °C) 1% osmium tetraoxide (OsO4) in the same buffer for 1 h, dehydrated in a graded series of acetone, embedded in Spurr’s epoxy resin and polymerized at 62 °C for 48 h. Ultrathin sections (60–90 nm in thickness) were cut on a Leica Ultracut UCT ultramicrotome (Leica, Wetzlar, Germany), placed on copper grids and stained sequentially with uranyl acetate and lead citrate according to Reynolds (1963). The sections were viewed under a JEOL 1010 transmission electron microscope (JEOL, Tokyo, Japan), equipped with a CCD digital camera Mega View III at 80 kV (Laboratory of Electron Microscopy, Institute of Parasitology, BC CAS).
Mega view 3
Manufactured by JEOL
Sourced in Japan
The Mega View III is a high-performance scanning electron microscope (SEM) from JEOL. It is equipped with advanced electron optics and imaging capabilities to provide high-resolution images of a wide range of samples. The Mega View III is designed for applications in materials science, nanotechnology, and other fields requiring detailed surface analysis.
Automatically generated - may contain errors
Lab products found in correlation
Jem 1010, by JEOL (3 mentions)
Jem 1200exiii, by JEOL (2 mentions)
Jem 1200ex 2, by JEOL (2 mentions)
1010 transmission electron microscope, by JEOL (1 mentions)
Ultracut uct ultramicrotome, by Leica (1 mentions)
Item uip software, by JEOL (1 mentions)
Nanosight ns500, by Malvern Panalytical (1 mentions)
Formvar coated copper grid, by Ted Pella (1 mentions)
Jem 1010 electron microscope, by JEOL (1 mentions)
Nano zsp, by Malvern Panalytical (1 mentions)
Collodion solution, by Merck Group (1 mentions)
Jem 1400 tem, by JEOL (1 mentions)
Ultra 45 diamond knife, by Electron Microscopy Sciences (1 mentions)
Durcupan acm resin, by Merck Group (1 mentions)
Jem 1010 80 kv electron microscope, by JEOL (1 mentions)
12 protocols using mega view 3
1
Ultrastructural Analysis of Flatworm D. latus
2
Nanocarrier Characterization by DLS and TEM
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Dynamic light scattering using a NICOMP 380 ZLS apparatus (Particle Sizing System, Menlo Park, CA, USA) with a 632.8 nm laser and a detection angle of 90° was utilized to evaluate the size and the size distribution of nanocarriers. After dilution in water (dilution factor 2), nanocarriers were analyzed. The water refractive index and viscosity values were 1.330 and 0.887 cP, respectively.
The ζ-potentials were evaluated by electrophoretic light scattering (NICOMP 380 ZLS apparatus, Particle Sizing System, Menlo Park, CA, USA), after the same sample dilution. Results were expressed as mean values ± standard deviation (SD) calculated from at least three independent samples.
The liposome morphology was evaluated by transmission electron microscopy (TEM; JEOL JEM-1200EXIII with TEM CCD camera Mega View III, Tokyo, Japan). Lipid nanocarriers, loaded on a copper grid, were stained with 1% (w/v) uranyl acetate (negative staining); measurement of NP sizes was carried out by ImageJ software (ImageJ 1.52a, NHI, Bethesda, MD, USA).
The ζ-potentials were evaluated by electrophoretic light scattering (NICOMP 380 ZLS apparatus, Particle Sizing System, Menlo Park, CA, USA), after the same sample dilution. Results were expressed as mean values ± standard deviation (SD) calculated from at least three independent samples.
The liposome morphology was evaluated by transmission electron microscopy (TEM; JEOL JEM-1200EXIII with TEM CCD camera Mega View III, Tokyo, Japan). Lipid nanocarriers, loaded on a copper grid, were stained with 1% (w/v) uranyl acetate (negative staining); measurement of NP sizes was carried out by ImageJ software (ImageJ 1.52a, NHI, Bethesda, MD, USA).
3
Cryo-TEM Characterization of mRNA Nanoparticles
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The ultrastructural morphology of the NPs and their mRNA nanocomplexes at optimum binding ratios (w/w) were determined by cryo-TEM, using a Jeol JEM-1010 transmission electron microscope containing a Soft Imaging System (SIS) fitted with a MegaView III digital camera with iTEM UIP software, operating at an acceleration voltage of 200 kV (Tokyo, Japan). The z-average hydrodynamic diameters and zeta (ζ) potentials were determined by nanoparticle tracking analysis (NTA, NanoSight NS500; Malvern Instruments, Worcestershire, UK) at 25 °C. NPs (1 mL) were diluted 1:100 in 18 MOhm and sonicated before analysis. Although the characterization of these NPs was reported previously by the authors [15 (link),16 (link)], the mRNA-based nanocomplexes are reported here for the first time.
4
Cryo-TEM Imaging of Cationic Liposomes
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The morphology of the cationic liposomes and lipoplexes (at optimal binding ratios) was examined by cryo-TEM. Liposomes or lipoplexes (2 µL) were deposited onto a formvar-coated copper grid (Ted Pella Inc., Redding, CA, USA) and contrasted 1:1 (v/v) with 4% saturated acidic uranyl acetate. The grids were plunged into liquid nitrogen at −180 °C using an injector system (Leica Microsystems EM CPC, Buffalo Grove, IL, USA), and the samples were examined using a JEOL JEM-1010 electron microscope (Jeol, Tokyo, Japan) operating at an accelerating voltage of 100 kV. The images were captured using the associated Soft Imaging System (SIS) MegaView III, bearing a side-mounted 3-megapixel digital camera.
5
Nanosystem Characterization Protocol
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Particle size (PS) and polydispersity index (PI) of all developed nanosystems were assessed at 25°C by photon correlation spectroscopy (PCS) at an angle of 90° (Beckman Coulter N5, Instrumentation Laboratory, Milan, Italy) after suitable dilution of samples with bidistilled and filtered water (0.22 µm, HA Millipore, Merck KGaA, Darmstadt, Germany).
Physical stability of nanoparticles was evaluated up to 90 days of storage at 2–8°C.
Zeta potential of the more promising formulation was measured at 25°C (Nano ZSP, Malvern Instruments, Alfatest, Rome, Italy).
These nanosystems were also analyzed by a transmission electron microscope (TEM; Jeol JEM-1200 EX II, Basiglio, Milan, Italy) equipped with a TEM CCD camera (Mega View III, Jeol, Basiglio, Italy), after sample deposition on copper grids (Formavar/Carbon 300 mesh, Agar Scientific, Assing, Rome, Italy).
Physical stability of nanoparticles was evaluated up to 90 days of storage at 2–8°C.
Zeta potential of the more promising formulation was measured at 25°C (Nano ZSP, Malvern Instruments, Alfatest, Rome, Italy).
These nanosystems were also analyzed by a transmission electron microscope (TEM; Jeol JEM-1200 EX II, Basiglio, Milan, Italy) equipped with a TEM CCD camera (Mega View III, Jeol, Basiglio, Italy), after sample deposition on copper grids (Formavar/Carbon 300 mesh, Agar Scientific, Assing, Rome, Italy).
6
Characterization of Scaffold Morphology and AgNPs
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Scaffold morphology was analyzed by means of SEM (Stereoscan 440 microscope, Leica Microsystems, Bensheim, Germany) after gold sputtering. The scaffolds were analyzed before and after the crosslinking procedure and after 1 week of hydration in distilled water. Nanofiber diameters were determined by an image analysis software (Image J, ICY, Institute Pasteur, Paris, France).
AgNPs loaded in the scaffolds were analyzed by transmission electron microscopy TEM (Transmission Electron Microscopy) (Jeol JEM-1200 EX II, Tokyo, Japan) equipped with TEM CCD camera Mega View III (Jeol, Tokyo, Japan). Nanofibers were directly electrospun onto copper grids.
FT-IR (Fourier Transform InfraRed) analysis was carried out by means of FTIR BX spectrum (Perkin Elmer, Italy). The infrared spectra were acquired in the range 4000–400 cm−1 and unloaded scaffolds and AgNPs loaded scaffolds were compared.
AgNPs loaded in the scaffolds were analyzed by transmission electron microscopy TEM (Transmission Electron Microscopy) (Jeol JEM-1200 EX II, Tokyo, Japan) equipped with TEM CCD camera Mega View III (Jeol, Tokyo, Japan). Nanofibers were directly electrospun onto copper grids.
FT-IR (Fourier Transform InfraRed) analysis was carried out by means of FTIR BX spectrum (Perkin Elmer, Italy). The infrared spectra were acquired in the range 4000–400 cm−1 and unloaded scaffolds and AgNPs loaded scaffolds were compared.
7
Transmission Electron Microscopic Analysis of SLN
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The morphological analysis of SLN was performed by means of a transmission electron microscope (TEM) Jeol JEM-1200n EXIII (Jeol USA Inc., Peabody, MA, USA), equipped with TEM CCD camera Mega View III. The images were captured after deposition of 10 μL of fresh SLN dispersion, properly diluted 1:2500 v/v in filtered deionized MilliQ water, on a grid (Nickel Square Mesh-Grid, 300 mesh, 3.05 mm diameter; Agar Scientific, Essex, UK) previously coated with Collodion solution (Sigma Aldrich, Milan, Italy).
8
Characterization of HA/CS Nanoparticles
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Physical properties of HA/CS NPs (mean diameter, PDI and surface charge) were assessed by dynamic light scattering (NICOMP 380 ZLS, Particles Sizing System, Santa Barbara, CA, USA). All the analyses were performed in triplicate for each formulation, and results were shown as mean ± SD. Morphological examination of NPs was performed by transmission electron microscopy (TEM) (JEOL JEM-1200EXIII with TEM CCD camera Mega View III, Tokyo, Japan) by using a negative staining (1% w/v uranyl acetate); NP sizes were determined by processing the TEM images by ImageJ software [50 (link)].
9
Transmission Electron Microscopy of M. smegmatis
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M. smegmatis strains were seeded at OD 0.001 in 7H9 with/without inducer and incubated at 37 °C with shaking for 30 hours. Bacterial cells were harvested and washed in PBS then fixed in 2.5% glutaraldehyde in PBS. An aliquot was saved for scanning electron microscopy (see below) and the remainder were pelleted in 3% agar, cooled, and cut into 1 mm3 cubes. Agar cubes were treated with 1% w/v aqueous osmium tetroxide, dehydrated through an ethanol series, and embedded in Spurr’s Resin. Samples were sectioned to a thickness of approximately 70 nm using a Reichert Ultracut E ultramicrotome and collected onto copper mesh grids. Dry grids were submerged briefly in methanol before staining in 2% (w/v) uranyl acetate for 30 minutes followed by Reynolds lead citrate for 5 minutes. Samples were viewed on a JEOL JEM-1400 TEM with an accelerating voltage of 100 kV. Images were collected using a Megaview III digital camera with iTEM software.
10
Cellular Uptake of La2O3 Nanoparticles
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To investigate the ability of La2O3NPs penetration into the CHANG and HuH-7 cells over 24 h were analyzed by transmission electron microscopy (TEM). The cells (3×104) were seeded in a cell culture dish (35 mm) for 24 hrs. The suspension of La2O3 NPs was exposed for 24 hrs. Shang et al17 (link) reported that uptake of nanoparticles was less at a higher concentration of nanoparticle exposure. So we selected a medium concentration (100 µg/mL) of La2O3 NPs for the uptake test. CHANG and HuH-7 cells were incubated with La2O3 NPs for 24 hrs in the same environmental condition.
After exposure, we have fixed the cells in 2.7% glutaraldehyde for 60 min and sections of cells were prepared according to Ciorîță et al.18 (link) The sections were placed on copper grids and examined with TEM Jeol JEM 1010 (JEOL, Tokyo, Japan), operated at 80 kV, and coupled with a Mega View III digital camera. To confirm if the electron-dense accumulations seen inside cells were La2O3 NPs.
After exposure, we have fixed the cells in 2.7% glutaraldehyde for 60 min and sections of cells were prepared according to Ciorîță et al.18 (link) The sections were placed on copper grids and examined with TEM Jeol JEM 1010 (JEOL, Tokyo, Japan), operated at 80 kV, and coupled with a Mega View III digital camera. To confirm if the electron-dense accumulations seen inside cells were La2O3 NPs.
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