Transmission Electron Microscopy (TEM) and Cryo-TEM analyses were performed using exosomes purified from cell culture supernatants by ultracentrifugation protocol [28 (link)]. Culture supernatants were centrifuged at 1000× g at room temperature for 10 min. The supernatants were collected and spun again at 10,000× g at room temperature for 30 min to remove cellular debris. The supernatants were filtered through 0.22 μm filters (Sigma-Aldrich, St. Louis, MO, USA), and exosomes were precipitated by ultracentrifugation at 100,000× g at room temperature for 2 h (Beckman Ultracentrifuge). The exosome pellet was resuspended in PBS for downstream analysis. Cryo-TEM was performed to demonstrate the purity and size of exosomes released from the HCV-infected cell culture using an FEI G2 F30 Tecnai TEM operating at 150 kV. The exosome samples were prepared on a lacey carbon-coated copper grid (200-mesh, electron microscopy sciences) using an automated plunging station (FEI Vitrobot). The sample solution was applied to the grid. The excess liquid was blotted by attached blotting papers for 2 s to produce a thin sample film that was immediately vitrified by plunging to liquid ethane. The grid with the sample cryogenically immobilized was transferred onto a single tilt cryo-specimen holder for imaging.
Tecnai g2 f30 tem
Manufactured by Thermo Fisher Scientific
The Tecnai G2 F30 TEM is a transmission electron microscope designed for high-resolution imaging and analysis of materials at the nanoscale. It features a field emission gun and advanced optics to provide exceptional image quality and resolution. The core function of this instrument is to enable detailed observation and characterization of the microstructure and composition of a wide range of materials.
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Lab products found in correlation
200 mesh lacey carbon grid, by Electron Microscopy Sciences (3 mentions)
Quanta 200 feg, by Thermo Fisher Scientific (2 mentions)
Ultracentrifuge, by Beckman Coulter (1 mentions)
Tecnai g2 f30 twin transmission electron microscope, by Thermo Fisher Scientific (1 mentions)
Lacey carbon coated copper grid, by Electron Microscopy Sciences (1 mentions)
Vitrobot, by Thermo Fisher Scientific (1 mentions)
S 4800 2 sem, by Hitachi (1 mentions)
D8 advance, by Bruker (1 mentions)
Spectrum 1000, by PerkinElmer (1 mentions)
Jsm 6510a, by JEOL (1 mentions)
Escalab 250xi xps spectrometer, by Thermo Fisher Scientific (1 mentions)
Labram hr evolution raman spectrometer, by Horiba (1 mentions)
Vitrobot mark 2, by Thermo Fisher Scientific (1 mentions)
Nova230, by Thermo Fisher Scientific (1 mentions)
Uv 2550, by Shimadzu (1 mentions)
Lambda 950 uv vis nir spectrometer, by PerkinElmer (1 mentions)
Quanta 200f microscope, by Thermo Fisher Scientific (1 mentions)
Dimension icon pt atomic force microscopy, by Bruker (1 mentions)
Labram hr800, by Horiba (1 mentions)
Tecnai g2 f30, by Thermo Fisher Scientific (1 mentions)
21 protocols using tecnai g2 f30 tem
1
Exosome Isolation and Cryo-TEM Analysis
2
Cryo-TEM Imaging of Extruded Liposomes
3
Cryo-TEM Imaging of Nanoparticle Complexes
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The morphology
of the complexes was characterized
by a FEI Tecnai G2 F30 twin transmission electron microscope operated
at 300 kV equipped with SDD EDS for elemental mapping. Cryo-TEM imaging
was done on an FEI G2 F30 Tecnai TEM operated at 150 kV. To prepare
the sample, a 200-mesh lacey carbon grid (Electron Microscopy Sciences)
was picked up with tweezers and mounted on the plunging station of
an FEI Vitrobot. Four microliters of the solution was applied to the
grid. The excess liquid was blotted by filter paper attached to arms
of the Vitrobot for 2 s to form a thin film. The sample was then vitrified
by plunging into liquid ethane. The vitrified sample was finally transferred
onto a single-tilt cryo specimen holder for imaging.
of the complexes was characterized
by a FEI Tecnai G2 F30 twin transmission electron microscope operated
at 300 kV equipped with SDD EDS for elemental mapping. Cryo-TEM imaging
was done on an FEI G2 F30 Tecnai TEM operated at 150 kV. To prepare
the sample, a 200-mesh lacey carbon grid (Electron Microscopy Sciences)
was picked up with tweezers and mounted on the plunging station of
an FEI Vitrobot. Four microliters of the solution was applied to the
grid. The excess liquid was blotted by filter paper attached to arms
of the Vitrobot for 2 s to form a thin film. The sample was then vitrified
by plunging into liquid ethane. The vitrified sample was finally transferred
onto a single-tilt cryo specimen holder for imaging.
4
Cryo-TEM Imaging of Vitrified Samples
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Cryo-TEM imaging
was done on a FEI G2 F30 Tecnai TEM operated at 200 kV. The sample
was prepared using a FEI Vitrobot. Five microliters of the solution
were applied to a 200-mesh lacey carbon grid (Electron Microscopy
Sciences). Excess liquid was blotted by filter paper attached to the
arms of the Vitrobot for 2 s to form a thin film. The sample was then
vitrified by plunging into liquid ethane followed by liquid nitrogen.
The vitrified sample was finally transferred onto a single tilt cryo-specimen
holder for imaging. The cryo-holder was maintained below 170 °C
to prevent sublimation of vitreous water. All size analyses on cryo-TEM
images were carried out using ImageJ software.
was done on a FEI G2 F30 Tecnai TEM operated at 200 kV. The sample
was prepared using a FEI Vitrobot. Five microliters of the solution
were applied to a 200-mesh lacey carbon grid (Electron Microscopy
Sciences). Excess liquid was blotted by filter paper attached to the
arms of the Vitrobot for 2 s to form a thin film. The sample was then
vitrified by plunging into liquid ethane followed by liquid nitrogen.
The vitrified sample was finally transferred onto a single tilt cryo-specimen
holder for imaging. The cryo-holder was maintained below 170 °C
to prevent sublimation of vitreous water. All size analyses on cryo-TEM
images were carried out using ImageJ software.
5
Cryo-TEM Imaging of Liposome-HCP Complexes
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Cryo-TEM imaging was conducted using an FEI G2 F30 Tecnai TEM operated
at 200 kV. The 5.0 mg/mL HCP and 2.5 mg/mL liposome aqueous solutions
prepared as described before were mixed in equal volume (v/v = 1:1)
and incubated at 25 °C overnight before imaging. 5 μL of
the sample solution was transferred to a 200-mesh lacey carbon-coated
copper grid (Electron Microscopy Sciences) mounted on the FEI Vitrobot
and blotted for 2 s to generate a sample thin film before plunging
into liquid ethane.
The morphology of HCPs and the liposome
complex was also analyzed using negative stained TEM. The 200-mesh
carbon-coated copper grid was pre-treated with a glow-discharger (LEICA
EM ACE 600) for 30 s to yield a negatively charged hydrophilic surface.
The sample solution (5 μL) was placed on the grid and left for
about 5 min before being blotted with a filter paper to remove excess
solution. 2 wt % uranyl acetate (5 μL) aqueous solution was
subsequently added on the grid and left for 20 s before being blotted
using a filter paper.
at 200 kV. The 5.0 mg/mL HCP and 2.5 mg/mL liposome aqueous solutions
prepared as described before were mixed in equal volume (v/v = 1:1)
and incubated at 25 °C overnight before imaging. 5 μL of
the sample solution was transferred to a 200-mesh lacey carbon-coated
copper grid (Electron Microscopy Sciences) mounted on the FEI Vitrobot
and blotted for 2 s to generate a sample thin film before plunging
into liquid ethane.
The morphology of HCPs and the liposome
complex was also analyzed using negative stained TEM. The 200-mesh
carbon-coated copper grid was pre-treated with a glow-discharger (LEICA
EM ACE 600) for 30 s to yield a negatively charged hydrophilic surface.
The sample solution (5 μL) was placed on the grid and left for
about 5 min before being blotted with a filter paper to remove excess
solution. 2 wt % uranyl acetate (5 μL) aqueous solution was
subsequently added on the grid and left for 20 s before being blotted
using a filter paper.
6
Cryo-TEM Imaging of Liposome-Peptide Complexes
7
Nanomaterial Characterization by Electron Microscopy
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Transmission electron microscopy (TEM), energy dispersive X-ray spectroscopy (EDS), and selected area electron diffraction (SAED) assays were performed using a FEI Tecnai G2 F30 TEM. Scanning electron microscopy (SEM) images were acquired using a S–4800II SEM (Hitachi, Japan). Dynamic light scattering and zeta potential were measured on a 90Plus PALS particle size and zeta potential analyzer. X-ray photoelectron spectroscopy (XPS) data were obtained using a 250Xi (Thermo Scientific, USA). X-ray diffraction (XRD) was performed using a D8 Advance (Bruker-AXS, Germany).
8
Characterization of PEI-based Nanocomposites
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FTIR analyses of PEIs and of B-BMSF@PEI samples before and after calcine were taken using a FTIR spectrometer (Spectrum 1000, Perkin Elmer, USA) at room temperature from the spectral region 400–4000 cm−1. Samples were thoroughly dried, crushed, and mixed with KBr. A Tecnai G2-F30 TEM instrument (FEI, The Netherlands) was used for evaluation of the shape and detailed morphology of B-BMSF@PEI. A JSM-6510A scanning electronic microscope (SEM) instrument (JEOL, Japan) was also employed to study the morphology and size (calculated from 200 nanoparticles). Meanwhile, the size distribution of B-BMSF@PEI was also evaluated using a Zeta-Potential/Particle Sizer (3000 HAS, UK) and the value was the average of three consecutive measurements. The porous structure and texture properties of B-BMSF@PEI were characterized by N2 desorption/adsorption tests using an adsorption analyzer (V-Sorb 2800P, Gold APP, China). Before analysis, sample was degassed at 120°C for 6 h. The specific surface area (SBET) of the prepared sample was analyzed by the Brunauer–Emmett–Teller (BET) method. The pore size distribution (WBJH) and pore volume (Vt) were derived from the adsorption branches of the isotherms using the Barrett-Joyner-Halenda (BJH) method.
9
Comprehensive Characterization of 3DG and GQDs/3DG Composites
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Transmission electron microscopy (TEM) images were acquired with a FEI Tecnai G2 F30 TEM. The samples were prepared by dispersing 3DG or GQDs/3DG composites in ethanol, then depositing them on copper mesh. Raman spectra were recorded on a HORIBA LabRAM HR Evolution Raman spectrometer with a 514 nm laser beam. Scanning electron microscopy (SEM) images were observed by a ZEISS MERLIN compact ultrahigh resolution Field Emission Scanning Electron Microcopy (FE-SEM). X-ray photoelectron spectroscopy (XPS) was recorded on a Thermo Fisher Scientific ESCALAB 250 Xi XPS spectrometer with Al Kα (1486.6 eV) as the X-ray source and a pass energy of 30 eV. The specific surface area of Brunauer-Emmett-Teller (BET) was measured by a Quantachrome Instruments QUADRASORB evoTM gas sorption surface area and pore size analyzer at ca. 77 K. Electrical conductivity was measured by an RTS-9 4-point probes resistivity measurement system from Guangzhou 4 Probes Tech. Co., Ltd (Guangdong, China). Electrochemical measurements were carried out on a CHI 660D electrochemical workstation under computer control.
10
Transmission Electron Microscopy of PA Gels
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An FEI Tecnai G2 F30 TEM instrument was employed. Overnight incubated PA gels were diluted to 50 µM. A total of 10 μL of diluted sample was placed onto the TEM grid and the sample was kept on the grid for 8 min. Then, the excess sample was removed from the surface by a micropipette. Negative staining was performed using 2% (w/v) uranyl acetate for 2 min incubation. Then, the grid was immediately washed with double-distilled water and left to dry.
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