Five microliters of the σNS–RNA complex, used in SV experiments (see above), was applied onto carbon-coated grids (Agar Scientific) to absorb for 1 min, after which the excess of sample was removed by blotting with filter paper. Grids were stained with 2% (w/v) uranyl acetate, and examined using JEOL 1200EX transmission microscope operating at 80 kV at 30 000× and 40 000× magnifications.
Carbon coated grids
Manufactured by Agar Scientific
Sourced in United Kingdom
Carbon-coated grids are a type of laboratory equipment used in electron microscopy. They provide a thin, uniform carbon film that is deposited onto a metal grid, typically made of copper. These grids serve as a support for specimens being examined under an electron microscope, allowing for high-resolution imaging and analysis.
Automatically generated - may contain errors
4 protocols using carbon coated grids
1
Negative Staining of σNS–RNA Complex
2
High-Resolution FESEM Mineralization Analysis
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HR FESEM was performed on NOVA NANOSEM-450, FEI (Hillsboro, OR, USA) with Zeiss lens. Aliquots from 96 well plate kept for mineralization in the presence of teeth protein extract were taken. Five microliters of each sample type were pipetted onto carbon-coated grids (Agar Scientific, Essex, UK). The grids were air-dried for several minutes. HR FESEM was operated at high voltage (HV) of 10.00 kV. Images obtained were from 100 nm to 10 μM and magnification ranging from 10,000× to 400,000×. Energy-dispersive X-ray (EDS or EDX) was also performed with the attached accessory within the same instrument. For EDS, HV of 20 kV was used.
3
Liposome Visualization via TEM
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Liposomes were placed on carbon-coated grids (Agar Scientific) and contrasted with 5% uranyl acetate solution. Electron microphotographs were obtained with a transmission electron microscope (JEM 1011; JEOL). Images were taken with an Orius SC1000 A charge-coupled device camera using image processing software (Gatan).
4
Structural Analysis of Octamer Protein
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3 µl of 20 µg/ml octamer was adsorbed onto carbon coated grids (Agar Scientific) glow discharged at 25mA for 25 s. The sample was stained with Uranyl Acetate and imaged using an FEI Tecnai12 Biotwin electron microscope operating at 120kV as described in (37 (link)). Micrographs were analysed using the EMAN2.0 (38 (link)). Particles were picked using a box size of 33, 33 and 35 nm giving final data sets of 6665, 10,108 and 1985 particles for wild-type, R141H and H207Q respectively, which was subject to iterative alignment and classification.
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