UV-vis absorption spectroscopy was performed on a Varian Cary 50 Scan UV-vis spectrophotometer. Photoluminescence (PL) spectra were measured by Horiba fluorescence spectrometer, in which the excitation wavelength was set as 350 nm. Transmission electron microscopy (TEM) graphs, scanning transmission electron microscopy (STEM) images were captured by a Talos transmission electron microscope (FEI Co.); prior to that, the CdSe NCs dissolved in hexane were dropped and dried on ultrathin carbon-supported copper grids. Fouriertransform infrared spectroscopy (FTIR) spectra were recorded by a Nicolet iS10 FT-IR Spectrometer (Thermo Scientific) equipped with a smart diamond attenuated total reflectance (ATR) accessory. Thermal gravimetric analysis (TGA) curve was measured by a TGA 4000 from PerkinElmer. The TGA data were obtained by heating the samples from 30°C to 600°C in the air atmosphere at a speed of 10°C/min. Powder X-ray diffraction (PXRD) measurement was performed on Empyrean with Cu LEF sealed tube and Pixcel1D detector from PANalytical.
Talos transmission electron microscope
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Talos transmission electron microscope is a high-performance instrument designed for advanced materials analysis and characterization. It provides high-resolution imaging, analytical capabilities, and versatile sample handling for a wide range of applications.
Automatically generated - may contain errors
Lab products found in correlation
Fei vitrobot mark 4, by Thermo Fisher Scientific (2 mentions)
Zetasizer pro, by Malvern Panalytical (2 mentions)
Fluorescence spectrometer, by Horiba (1 mentions)
Tga 4000, by PerkinElmer (1 mentions)
Pixcel 1d detector, by Malvern Panalytical (1 mentions)
Nicolet is10 ft ir spectrometer, by Thermo Fisher Scientific (1 mentions)
Cary 50 scan uv vis spectrophotometer, by Agilent Technologies (1 mentions)
Beckman 70.1 ti rotor, by Beckman Coulter (1 mentions)
Whatman, by Cytiva (1 mentions)
Ceta 16m detector, by Thermo Fisher Scientific (1 mentions)
Cf400 cu grid, by Electron Microscopy Sciences (1 mentions)
Superose 6 10 300 column, by GE Healthcare (1 mentions)
Solarus 950, by Ametek (1 mentions)
Tia v4, by Thermo Fisher Scientific (1 mentions)
Ivis spectrum imaging system, by PerkinElmer (1 mentions)
Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (1 mentions)
Falcon 3 detector, by Thermo Fisher Scientific (1 mentions)
Epu 2, by Thermo Fisher Scientific (1 mentions)
Titan krios, by Ametek (1 mentions)
Apreo 2 s lovac scanning electron microscope, by Thermo Fisher Scientific (1 mentions)
12 protocols using talos transmission electron microscope
1
Comprehensive Characterization of CdSe Nanocrystals
2
Protein Staining and TEM Imaging
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Protein from the final step of this procedure was stained using 3% Uranyl Acetate on a 400-mech continuous carbon grid. Images were acquired using an FEI Talos transmission electron microscope operating at 200 kV, with 1.25 s exposures, a dose rate of 19 e- Å-2, and a nominal magnification of x57000.
3
Fecal Virus Concentration for DEM
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Two fecal samples from 1 animal (case 2) were pooled for direct electron microscopy (DEM). Four mL of fecal sample was diluted with water to a final volume of 13 mL and clarified by centrifuging at 2400 x g for 20 minutes at 30 C. The supernatant was filtered sequentially through 5.0, 0.8, 0.45, and 0.2 micron syringe filters. Seven milliliters of the filtered supernatant were placed in a 13.5 mL Beckman polycarbonate thick wall centrifuge tube (Beckman Instruments, Part number 355630) and centrifuged at 55 000 rpm in a Beckman 70.1 Ti rotor for 45 minutes at 5 C. The protein pellet on the bottom of the centrifuge tube was suspended for approximately 20-30 minutes in 1.0 mL water and placed into a 1.5 mL labeled freezing vial. The pellet suspension was mixed with 2% neutralized phosphotungstic acid (PTA) at an approximate ratio of 1:10 to form a slurry that was applied to a carbon coated Formvar grid and excess slurry was wicked away with Whatman #1 filter paper wedges. Grids were examined on an FEI Talos transmission electron microscope at 80 KeV accelerating voltage (FEI, Hillsboro, OR).
4
Structural Analysis of Deglycosylated gB-Fab Complex
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The postfusion gB was deglycosylated by digestion with Endo H (10% w/w) for 12 hours at 4°C and purified using a Superose6 10/300 column. Deglycosylated gB was mixed with a molar excess of 3–25 Fab and run over a Superose6 10/300 column (GE Healthcare) using 2 mM Tris pH 8.0, 200 mM NaCl, 0.02% NaN3. A CF400-Cu grid (Electron Microscopy Sciences) was plasma cleaned for 30 seconds in a Solarus 950 (Gatan) using a 4:1 mixture of O2 to H2. The purified complex was diluted to a concentration of 0.025 mg/mL and mixed with additional 3–25 Fab to fully saturate the gB trimer before being deposited onto the prepared grid. Micrographs were collected using a 200 kV Talos transmission electron microscope (Thermo Fisher) equipped with a Ceta 16M detector (Thermo Fisher) at a magnification of 92,000×, corresponding to a calibrated pixel size of 1.63 Å. Data were collected manually using TIA v4.14 (Thermo Fisher) at a defocus range of 1.8 to 3.1 μm. CTF-estimation, particle picking and preliminary 2D classification were performed using cisTEM [63 (link)] before particles were exported into cryoSPARC v2.9.0 [64 (link)] for ab initio model generation and 3D classification.
5
Characterizing Lipid Nanoparticle Formulations
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LPPs were diluted into PBS for determining size distribution and zeta potential. Zeta potentials were measured by diluting LPPs into folded capillary zeta cell at pH 4.3 or pH 7.2 and loaded into Zetasizer Pro (Malvern Panalytical, UK). Particle size distribution was measured with dynamic light scattering. For cryo-electron microscopy, LPPs were transferred onto a glow-discharged ultrathin carbon-coated copper grid, blotted for 2 s with filter paper in FEI Vitrobot Mark IV (Thermo Fisher Scientific, USA), followed by quick plugging into liquid ethane. Frozen grids were loaded into a Talos transmission electron microscope (Thermo Fisher Scientific, USA) equipped with a field emission gun operated at 200 kV. Images were recorded on a direct electron detector (ED20). For in vivo imaging, mRNAs encoding luciferase were encapsulated by LNP or LPP and injected in both legs of BALB/c mice (10 μg/mouse). Bioluminescence image of mice was captured using IVIS Spectrum Imaging System (PerkinElmer, USA) at 6, 12, 24 and 48 h post-injection.
6
Transmission Electron Microscopy of sEVs
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The sEV pellets were resuspended in 50 μL of DPBS (Gibco, Billings, MT, USA) and then prepared for microscopy, using 15 μL of the sEV pellets and a 5-fold dilution in double-distilled H2O. The samples were incubated for 1 min in ozone pre-treated copper grids, then fixed with Uranyl acetate for 1 min, and then dried in a 60 °C oven for 4 min. After that, the samples were visualized in the microscope, and at least 5 photos per condition were captured. The qualitative morphology assessment of the sEVs was analyzed using the Talos transmission electron microscope (Thermo Scientific, Waltham, MA, USA).
7
LNP Characterization via DLS and Cryo-EM
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For determining the particle sizes and zeta potential of LNPs, LNPs were diluted into deionized water at pH 4.0 or pH 7.4. Diluted LNPs were added into folded capillary zeta cells and loaded into Zetasizer Pro (Malvern Panalytical). Particle size distribution and polydispersity index were measured with dynamic light scattering. Zeta potential was measured by Zeta. For cryo-electron microscopy, LNPs were transferred onto a glow-discharged ultrathin carbon-coated copper grid, blotted for 2 s with filter paper in FEI Vitrobot Mark IV (Thermo Fisher), followed by quick plugging into liquid ethane. Frozen grids were loaded into a Talos transmission electron microscope (Thermo Fisher Scientific) equipped with a field emission gun operated at 200 kV. Images were recorded on a direct electron detector (ED20).
8
Cryo-EM Data Acquisition with Titan Krios
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Cryo-EM grids were first checked on a Talos transmission electron microscope equipped with a Falcon III detector (Thermo Fisher Scientific) operated in linear mode. The images were recorded at a nominal magnification of 120,000x, corresponding to a pixel size of 0.86 Å/pixel, with a defocus setting of -3.0 μm. Suitable cryo-EM grids were recovered for further data collection on a 300 kV Titan Krios transmission electron microscope hosting a K3 detector (with GIF Bio-Quantum Energy Filters, Gatan) operating in super-resolution mode and using EPU-2.7.0 software (Thermo Fisher Scientific). The raw movie stacks were recorded at a magnification of 105,000×, corresponding to a pixel size of 0.83 Å/pixel (super-resolution 0.415 Å/pixel). The defocus range was set to −1.5 to -2.5 μm and the slit width of energy filters was set to 20 eV. Forty frames of non-gain-normalized tiff stacks were recorded with a dose rate of ~16 e-/Å2 per second and the total exposure time was set to 2.5 s, resulting in an accumulated dose of ~40 e-/Å2 (~1.0 e-/Å2 per frame). The parameters for cryo-EM data acquisition are summarized in Table 1 .
9
TEM Visualization of Protein Samples
10
Analyzing Autophagosome Formation by TEM
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Ultrathin sections of cells for electron microscopic analysis were prepared at the Neuroscience Translational Research Solution Center at Dong-A University. Ultrathin sections were observed and photographed using a Talos transmission electron microscope (Thermo Fisher Scientific, Waltham, MA, USA) or an Apreo 2 S LoVac scanning electron microscope (Thermo Fisher Scientific) at the Neuroscience Translational Research Solution Center, Dong-A University. To analyze autophagosome formation upon CCCP or DFP treatment, HEK293 cells treated with CCCP or DFP were analyzed via transmission electron microscopy. At least ten cells per group were examined to determine the number of autophagosomes, and the results are presented as the mean ± SD.
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