HeLa cells on coverslips were infected at m.o.i. 10 with WT VACV in the presence of AraC for 6 h then shifted into the indicated CGs. At 20 h post-shift, the coverslips were fixed in EM-grade 2 % paraformaldehyde/2 % glutaraldehyde (TAAB Laboratories Equipment) in 0.1 M sodium cacodylate, secondarily fixed for 1 h in 1 % osmium tetraoxide/1.5 % potassium ferricyanide at 4o C and then treated with 1 % tannic acid in 0.1 M sodium cacodylate for 45 min at room temperature. Samples were then dehydrated in sequentially increasing concentration of ethanol solutions and embedded in Epon resin. Coverslips were inverted onto prepolymerised Epon stubs and polymerized by baking at 60oC overnight. The 70 nm thin sections were cut with a Diatome 45 diamond knife using an ultramicrotome (UC7; Leica). Sections were collected on 1×2 mm formvar-coated slot grids and stained with Reynolds lead citrate. All samples were imaged using a transmission electron microscope (Tecnai T12; FEI) equipped with a charge-coupled device camera (SIS Morada; Olympus).
Sis morada
Manufactured by Olympus
Sourced in Germany
The SIS Morada is a high-performance scientific imaging camera designed for advanced microscopy applications. It features a large sensor, high-resolution imaging, and low-noise performance to capture detailed, high-quality images for research and analysis purposes.
Automatically generated - may contain errors
Lab products found in correlation
Tecnai t12, by Thermo Fisher Scientific (3 mentions)
Jem 1010, by JEOL (1 mentions)
Tecnai g2 spirit, by Thermo Fisher Scientific (1 mentions)
Glutaraldehyde, by TAAB (1 mentions)
Vt1200, by Leica (1 mentions)
Fei morgagni 268d, by Thermo Fisher Scientific (1 mentions)
Uranyl acetate, by Merck Group (1 mentions)
Uct ultramicrotome, by Leica (1 mentions)
7 protocols using sis morada
1
Ultrastructural Analysis of VACV Infected Cells
2
Characterization of drug-loaded nanoparticles by TEM
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The morphology of RA-loaded PLGA NPs and PC NPs was assessed by transmission electron microscopy (TEM). Briefly, 5 μL of the sample was adhered onto glow discharged carbon-coated grids for 60 s. The remaining liquid was removed using a cellulose filter paper and was stained with 2 wt% phosphotungstic acid for 60 s. A Jeol JEM1010 transmission electron microscope (Tokyo, Japan) was used to visualize the samples using an accelerating voltage of 200 KeV in a bright-field image mode. Digital images were acquired using a digital camera (Olympus SIS Morada).
3
Electron Microscopy of Viral Fractions
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LB-Core and Core samples were prepared as described (see Viral Fractionation) and prepared for EM by fixation in 1.5% glutaraldehyde / 2% EM-grade paraformaldehyde (TAAB) in 0.1M sodium cacodylate for 45 min at RT. Briefly, samples were treated with reduced osmium, tannic acid, dehydrated through an ethanol series and embedded in epon resin [96 (link)]. Sections were collected on formvar coated slot grids, stained with lead citrate and imaged using a transmission electron microscope (Tecnai T12, Thermo Fisher Scientific) equipped with a charge-coupled device camera (SIS Morada; Olympus).
4
TEM Imaging of Transfected HUVECs
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HUVECs were transfected with WT or Quad-K JAM-C–HRPout constructs and plated onto glass coverslips. The coverslips were fixed in EM-grade 2% paraformaldehyde, 1.5% glutaraldehyde (TAAB Laboratories Equipment, Ltd., Aldermaston, UK) and washed in 0.05 M Tris HCl (pH 7.6) before incubation with 0.075% DAB, 0.02% hydrogen peroxide for 30 min in the dark. The coverslips were washed in 0.1 M sodium cacodylate and secondarily fixed in 1% osmium tetraoxide, 1.5% potassium ferricyanide and then treated with 1% tannic acid. Samples were then dehydrated and embedded in Epon resin. Coverslips were inverted onto prepolymerised Epon stubs and polymerised by baking at 60°C overnight. Seventy-nm–thick sections were cut with a diatome 45° diamond knife using an ultramicrotome (UC7; Leica, Wetzlar, Germany). Sections were collected on 1 × 2 mm Formvar-coated slot grids and stained with Reynolds lead citrate. Samples were imaged using a transmission electron microscope (Tecnai G2 Spirit; FEI, Thermo Fisher Scientific) and a charge-coupled device camera (SIS Morada; Olympus).
5
Ultrastructural Analysis of Neuronal Tissue
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LNs were fixed overnight in 2% PFA/1.5% glutaraldehyde (both EM grade from TAAB) in 0.1M sodium Cacodylate at 4°C and embedded in 2.8% low melting point agarose dissolved in PBS. Slices of 100 μm thickness were then cut in cold PBS using a vibrating microtome (VT1200S; Leica) and returned to fresh fix solution for a further 15 mins. Slices were then secondarily fixed for 1 h in 1% osmium tetraoxide/1.5% potassium ferricyanide at 4°C and then treated with 1% tannic acid in 0.1M sodium cacodylate for 45min at room temperature. Samples were then dehydrated in sequentially increasing concentration of ethanol solutions, and embedded in Epon resin. The 70nm ultrathin resin sections were cut with a Diatome 45° diamond knife using an ultramicrotome (UC7; Leica). Sections were collected on 1 × 2mm formvar-coated slot grids and stained with Reynolds lead citrate. All samples were imaged using a transmission electron microscope (Tecnai T12; FEI) equipped with a charge-coupled device camera (SIS Morada; Olympus).
6
Immunogold Labeling of Cellulase 7
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Ultrathin sections on grids was pre-incubated in 10 mM PBS for 10 min. Afterwards, unspecific binding sites were blocked by incubation in 10 mM PBS supplemented with 2% (w/v) BSA for 45 min. Thereafter, grids were treated with primary rabbit anticellulase 7 antibody (anti-Sl-CEL7; Sigma-Genosys) diluted 1:50 with 10 mM PBS supplemented with 2% (w/v) BSA for 1 h at RT. Then, sections were washed in 10 mM PBS with 0.05% (v/v) Tween20 four times for 10 min and once in 10 mM PBS for 10 min. They were incubated for 1 h at RT in secondary goat anti-rabbit IgG conjugated with 15 nm colloidal gold particles (British BioCell International, Cardiff, UK) diluted 1:50 with 10 mM PBS. They were washed three times in 10 mM PBS and rinsed twice in distilled water. Grids were counterstained with 2% (w/v) aqueous solution of uranyl acetate (Sigma-Aldrich) for 10 min and washed in distilled water
five times for 2 min. In negative controls, primary antibody was omitted. Grids were examined in an FEI 268D "Morgagni" (FEI Company, Hillsboro, OR, USA) transmission electron microscope operating at 80 kV. Images were taken with an SIS "Morada" (Olympus-SIS, Münster, Germany) digital camera.
five times for 2 min. In negative controls, primary antibody was omitted. Grids were examined in an FEI 268D "Morgagni" (FEI Company, Hillsboro, OR, USA) transmission electron microscope operating at 80 kV. Images were taken with an SIS "Morada" (Olympus-SIS, Münster, Germany) digital camera.
7
Arabidopsis-Heterodera schachtii Pathosystem
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Arabidopsis plants were grown and inoculated with H. schachtii as described above. Samples of uninfected roots and roots containing syncytia were collected from wild-type (Col0) and lsd1 mutant plants at 7 and 12 dpi, and processed for light and transmission electron microscopy examinations, as described by Golinowski et al. (1996) (link) and Sobczak et al. (1997) (link). Sections were cut using a Leica UCT ultramicrotome, stained with uranyl acetate and lead citrate, and examined using a FEI 268 D ‘Morgagni’ (FEI Comp., Hillsboro, OR, United States) transmission electron microscope equipped with an SIS ‘Morada’ (Olympus SIS, Münster, Germany) digital camera.
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