The sectioned lizard tails were fixed in 4% paraformaldehyde and 2% glutaraldehyde in 0.1M cacodylate buffer at pH 7.4 (23 , 32 (link)). The tails were fixed for 12 to 16 h at room temperature and then rinsed four times in cacodylate buffer, then postfixed and stained with 1% osmium tetroxide and 1.5% potassium ferricyanide in 0.1M cacodylate buffer for 1 h. Tissues were then washed four times in cacodylate, dehydrated in increasing concentrations of ethanol, rinsed twice in propylene oxide, infiltrated with increasing concentrations of Agar 100 resin in propylene oxide, and embedded in fresh resin. Ultra-thin (80 nm) cross-sections were cut with a diamond knife on a LKB 3 microtome. The cross-sections were then placed on carbon-formvar-coated copper grids and were poststained with uranyl acetate and lead citrate. TEM imaging on the ultra-thin biological tissues was performed using a Tecnai 12 TEM 100 kV (Phillips) equipped with MegaView II CCD camera. The TEM images were collected from three hatchling specimens and two adult specimens, which include hundreds of cells from hatchling to adult in total.
Megaview 2 ccd camera
Manufactured by Philips
The MegaView II CCD camera is a high-performance imaging device designed for scientific and industrial applications. It features a charged-coupled device (CCD) sensor that captures detailed images and data. The camera is capable of capturing high-resolution images and video, making it suitable for various imaging-based research and analysis tasks.
Automatically generated - may contain errors
Lab products found in correlation
Morgagni 268d, by Philips (2 mentions)
Tecnai 12 tem 100 kv, by Philips (1 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions)
Osmium tetroxide, by Merck Group (1 mentions)
Propylene oxide, by Merck Group (1 mentions)
Agar 100 resin, by Agar Scientific (1 mentions)
Technai 12, by Philips (1 mentions)
Normal goat serum (ngs), by Thermo Fisher Scientific (1 mentions)
Lr white resin, by Merck Group (1 mentions)
Tecnai 12, by Philips (1 mentions)
Ps nh2, by Bangs Laboratories (1 mentions)
6 protocols using megaview 2 ccd camera
1
Ultrastructural Analysis of Lizard Tail Regeneration
2
Ultrastructural Analysis of Olive Leaves
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We analyzed olive leaves of both varieties taken at 3 selected time points (T0, T4, and T8). The protocol is detailed in Behr et al. [64 (link)]. For transmission electron microscopy (TEM), samples were fixed in 3% glutaraldehyde in cacodylate buffer (0.066 M, pH 7.2), for 1 h at room temperature. After fixation, samples were rinsed with cacodylate buffer and post-fixed with osmium tetroxide 1% in cacodylate buffer for 1 h. Then, samples were rinsed with water and dehydrated gradually in increasing concentrations of ethanol (from 10% to 100%). Samples were embedded in Spurr’s resin [65 (link)], polymerized for 8 h at 70 °C, and then cut into 600-Å sections using an LKB Nova ultramicrotome provided with diamond knife. Sections were stained with uranyl acetate and lead citrate for 10 min, respectively, and finally observed with a Philips Morgagni 268D transmission electron microscope operating at 80KV and equipped with a MegaView II CCd camera (Philips Electronics). Three different sets of experiments were subjected to TEM analysis.
3
Ultrastructural Analysis of Pancreatic Mitochondria
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Pancreas was fixed with 4% paraformaldehyde and 2.5% glutaraldehyde (Electron Microscopy Sciences), post-fixed with 1% osmium tetroxide (Sigma), and dehydrated with increasing concentrations of ethanol followed by propylene oxide (Sigma). For embedding we used Agar 100 Resin (Agar Scientific). For imaging we used 80-nm sections stained with 5% uranyl acetate for 10 min followed by 10 min with lead citrate. Samples were visualized with a transmission electron microscope (Technai 12, Phillips) equipped with a MegaView II CCD camera. To assess structural defects in mitochondria, we determined for each EM-imaged mitochondrion whether it was swollen or had defective cristae. Docked granules were counted up to 200 nm from the plasma membrane and calculated as number of granules per membrane length.
4
Immunogold Labeling of ALIX in Pollen
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For electron microscopy, the transmission electron microscope (TEM) Philips Morgagni 268 D set at 80 kV was employed, and images were captured with a MegaView II CCD Camera (Philips Electronics, The Netherlands) and analyzed with the microscope software (AnaliSYS). Immunogold labeling was carried out following literature (Parrotta et al., 2019 (link)). Briefly, germinated pollen was dehydrated in growing concentrations of ethanol, and then infiltrated with LR white resin (Sigma-Aldrich, Italy). Thus, the resin was encapsulated and polymerized in an oven at 40°C for 2 days. The resin was then sectioned, and the sections were blocked in 5% normal goat serum (Invitrogen, Italy) for 20 min and then incubated in a 1:50 dilution of the anti-ALIX antibody for 1 h. Three sections were selected as negative controls and they were not incubated with the primary antibody. Finally, the excess of primary antibody was washed in 50 mM Tris–HCl pH 7.6, 0.9% NaCl, 0.1% Tween, and all the sections were incubated for 45 min with a dilution 1:20 of goat anti-rabbit secondary antibody conjugated with 15 nm gold particles (BioCell, Italy). Sections were washed with distilled water and counterstained first with 2% uranyl acetate for 10 min, and then with lead citrate for 5 min. No osmium was employed in the preparation of the samples. At least 50 pollen tubes and grains were analyzed per sample.
5
Tissue Preparation for TEM Imaging
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Samples for transmission electron microscopy (TEM) were fixed with a mixture of 2.5% glutaraldehyde in 0.1 M phosphate buffer, pH 7.2, for 2 h and then washed with 0.1 M phosphate buffer. After osmification, dehydration and embedding (Epon), the tissue was sectioned using an LKB-ultrotome 8800 III. Thick sections (1 μm) were stained with toluidine blue for light microscopy and thin sections (70 nm) were stained with uranyl acetate and Reynold’s lead citrate for transmission electron microscopy (TEM) and observed with a Tecnai 12 (Phillips, Eindhoven, The Netherlands) TEM equipped with MegaView II CCD camera and AnalySIS version 3.0 software.
6
Characterization of 50 nm Amino Polystyrene Nanoparticles
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Primary characterization of unlabelled 50 nm amino polystyrene NPs (PS-NH2), purchased from Bangs Laboratories, was performed as described in our previous paper (Della Torre et al., 2014) . Primary particle diameter of PS-NH2 was determined by transmission electron microscopy (Philips Morgagni 268D electronics, at 80 KV and equipped with a MegaView II CCd camera. Artificial sea water (ASW) was prepared according to ASTM protocol (pH 8, salinity 36 ‰) (ASTM 2004) and filtered with 0.22 µm membrane. PS-NH2 suspensions (50 µg/ml) were prepared in ASW, quickly vortexed prior to use but not sonicated. Size (Z-average and polydispersity index, PDI) and zeta potential (ζ-potential, mV) were determined by Dynamic Light Scattering (Malvern instruments), using a Zetasizer Nano Series software, version 7.02 (Particular Sciences, UK). Measurements were performed in triplicate, each containing 11 runs of 10 seconds for determining Z-average, 20 runs for the ζ-potential.
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