The morphology of the prepared KLA-5-FU/PTX Lps was analyzed using cryo-transmission electron microscopy (FEI Titan Krios Transmission Electron Microscope, Thermo Scientific, MA, USA).
Fei titan krios transmission electron microscope
Manufactured by Thermo Fisher Scientific
Sourced in United States
The FEI Titan Krios transmission electron microscope is a high-performance, cryogenic electron microscope designed for advanced structural biology research. It provides exceptional image quality and resolution, enabling detailed analysis of biological samples at the atomic level.
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Lab products found in correlation
Vitrobot mark 4, by Thermo Fisher Scientific (2 mentions)
Vitrobot plunge freezer, by Thermo Fisher Scientific (1 mentions)
Pelco easiglow device, by Ted Pella (1 mentions)
Solarus plasma cleaner, by Ametek (1 mentions)
Vitrobot mark 4 instrument, by Thermo Fisher Scientific (1 mentions)
K2 direct electron detector, by Ametek (1 mentions)
Solarus 950 plasma cleaner, by Ametek (1 mentions)
5 protocols using fei titan krios transmission electron microscope
1
Cryo-TEM Analysis of KLA-5-FU/PTX Liposomes
2
Cryo-EM Sample Preparation Protocol
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The sample was diluted to a concentration of 0.25 mg/mL. For cryo-EM sample preparation, 4.0 μL of the purified complex were applied to glow discharged Quantifoil 2/1 grids, blotted for 3.5 s with force ‘4’ in a Vitrobot Mark IV (Thermo Fisher) at 100% humidity and 4°C, and plunge vitrified in liquid ethane cooled by liquid nitrogen.
Electron micrographs were acquired with a FEI Titan Krios transmission electron microscope (ThermoFisher) using SerialEM software.54 (link) Movie frames were recorded at a nominal magnification of 81,000× (calibrated physical pixel size: 1.06Å/px) using a K2 direct electron detector (Gatan) and a GIF quantum energy filter (Gatan) at 20 eV slit width. The total electron dose of approximately 47 electrons per Å2 was distributed over 35 frames and 3281 micrographs were collected.
Electron micrographs were acquired with a FEI Titan Krios transmission electron microscope (ThermoFisher) using SerialEM software.54 (link) Movie frames were recorded at a nominal magnification of 81,000× (calibrated physical pixel size: 1.06Å/px) using a K2 direct electron detector (Gatan) and a GIF quantum energy filter (Gatan) at 20 eV slit width. The total electron dose of approximately 47 electrons per Å2 was distributed over 35 frames and 3281 micrographs were collected.
3
Vitrified Exosome Imaging Protocol
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Purified exosomes in PBS buffer were vitrified on Quantifoil Cu R200 2/2 (Electron Microscopy Sciences, Ref. No.: Q2100CR2) grids. Prior to sample application, the grids were glow discharged using a Pelco easiGlow device (Ted Pella Inc.) at 15 mA for 30 s. Samples were deposited by transferring 3 μl of sample onto the glow-discharged side of the grid, blotted, then plunge frozen in liquid ethane, using a Vitrobot plunge freezer (Thermo Fisher Scientific), with the following settings: 22°C, 80% humidity, Blot-force = –5, 60 s wait time and a blotting time of 3 s. All data were collected on an FEI Titan Krios transmission electron microscope (Thermo Fisher Scientific) operated at 300 keV and equipped with a Gatan BioQuantum energy filter and a K2 direct electron detector. A condenser aperture of 70 μm and no objective aperture were chosen for data collection. Data were acquired in parallel illumination mode using EPU (Thermo Fisher Scientific) software at a nominal magnification of 33 kx (4.3 Å pixel size).
4
Cryo-EM Protocol for Structural Elucidation
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A droplet (2.5 μL) of SOS1 protein sample was applied to the holey carbon grids (Cu R1.2/1.3 300 mesh, Quantifoil), which were glow-discharged beforehand in H2-O2 condition for 1 min using a Solarus plasma cleaner (Gatan, USA). The grids were then blotted with a Vitrobot Mark IV (Thermo Fisher Scientific, USA) for 4.5 s under 4 °C and 100% humidity and flash-frozen in liquid ethane cooled by liquid nitrogen. Cryo-EM data were collected on a 300-kV FEI Titan Krios transmission electron microscope (Thermo Fisher Scientific, USA) equipped with a Gatan K2 Summit direct electron detector and a GIF quantum energy filter. The SerialEM45 (link) was used to collect movie stacks automatically at a magnification of 130,000×, which corresponds to a pixel size of 0.52 Å across a defocus range of –1.2 μm to –2.2 μm. Each movie stack was dose-fractionated in 32 frames with a total dose of 60 e–/Å2. The dose rate was set to 9.0 e–/pixel/s.
5
Cryo-EM Protocol for PEDV Spike Protein
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CF-2/2 grids (Electron Microscopy Sciences) were plasma cleaned for 30 s in a Gatan Solarus 950 plasma cleaner with a 4:1 O2/H2 ratio. PEDV S CV777 at a concentration of 0.4 mg/ml was mixed with 2 mM Tris (pH 8.0), 200 mM NaCl, 0.02% NaN3, and 0.01% Amphipol A8-35 and was deposited onto the grids before being blotted for 6 s and plunge-frozen in liquid ethane using a Vitrobot Mark IV instrument (Thermo Scientific). The frozen grid was imaged using an FEI Titan Krios transmission electron microscope (Thermo Scientific) equipped with a K2 direct electron detector (Gatan) at a nominal magnification of ×22,500, corresponding to a calibrated pixel size of 1.075 Å/pixel. A total of 3,186 movies were collected using Leginon (45 (link)). A full description of the cryo-EM data collection parameters can be found in Table 1 .
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