Thin sections (100 nm) obtained from the block face were imaged at 120 kV on a FEI-Tecnai™ Spirit TEM system for assessment of tissue contrast and fixation, and for comparison with FIB-SEM images. Images were collected as 2048 × 2048 pixel, 16-bit gray scale using the FEI’s TEM Imaging & Analysis (TIA) interface on an Eagle™ 2K CCD multiscan camera.
Tecnai spirit tem system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Tecnai™ Spirit TEM system is a transmission electron microscope (TEM) designed for high-resolution imaging and analysis of samples at the nanoscale level. The system provides advanced capabilities for researchers and scientists in fields such as materials science, nanotechnology, and life sciences.
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Lab products found in correlation
9 protocols using tecnai spirit tem system
1
Ultrastructural Imaging of Biological Samples
2
Exosome Isolation and Characterization
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Stromal cell exosomes were isolated by sucrose step-gradient then washed in 0.22 μm filtered PBS. 10 μl was deposited onto glow discharged carbon formvar 400 mesh copper grids (Ted Pella 01822 F) for 3 min, rinsed 15 secs in water, wicked on Whatman filter paper 1, stained for 45 secs in filtered 1.33% (w/v) uranyl acetate, wicked and air dried. Samples were imaged at 120kV on a FEI Tecnai Spirit TEM system. Images were acquired as 2048 × 2048 pixel, 16-bit gray scale files using the FEI’s TEM Imaging and Analysis (TIA) interface on an Eagle 2K CCD multiscan camera.
3
TEM Analysis of Purified Phage Lysate
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PDX was analysed using transmission electron microscopy (TEM). Purified high-titre lysate was precipitated in 25 % PEG 6000–8000 in 2.5M NaCl and stored overnight at 4 ˚C. Precipitated phages in solution were centrifuged at 4 ˚C, 4000g for 10 min. The supernatant was removed and the pellet was suspended in 40 µl of 50 mM Tris, pH 7.4, 10 mM MgSO4. The phage suspension was placed on copper-coated formvar grids (Ted Pella, Inc., Redding, CA, USA) and negatively stained with 1 % uranyl acetate. Samples were examined under a FEI Tecnai Spirit TEM system at an operating voltage of 120 kV (FEI, Hillsboro, OR, USA). All TEM was performed at the Multi-Scale Microscopy Core (MMC) with technical support from the Oregon Health and Science University (OHSU)/FEI Living Lab and the OHSU Center for Spatial Systems Biomedicine (OCSSB).
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PDX was analysed using transmission electron microscopy (TEM). Purified high-titre lysate was precipitated in 25 % PEG 6000–8000 in 2.5M NaCl and stored overnight at 4 ˚C. Precipitated phages in solution were centrifuged at 4 ˚C, 4000
4
Negative Staining of Exosomes
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10 μl of Exosome preparations were deposited onto glow discharged carbon formvar 400 Mesh copper grids (Ted Pella 01822-F) for 3 min, rinsed 15 secs in water, wicked on Whatman filter paper 1, stained for 45 secs in filtered 1.33% (w/v) uranyl acetate, wicked and air dried. Samples were imaged at 120 kV on a FEI Tecnai™ Spirit TEM system. Images were acquired as 2048 × 2048 pixel, 16-bit gray scale files using the FEI’s TEM Imaging & Analysis (TIA) interface on an Eagle™ 2K CCD multiscan camera.
5
Visualizing HBV Virions by Electron Microscopy
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Supernatants from rhesus macaque PH, confirmed for infection with HBV (Fig. 2c ), were diluted 1:10 in 4% paraformaldehyde. 10 µl of the fixed viral suspensions were deposited onto glow discharged carbon formvar 400 Mesh copper grids (Ted Pella 01822-F) for 3 min, rinsed 15 s in water, wicked on Whatman filter paper 1, stained for 60 s in filtered 1.33 % (w/v) uranyl acetate in water, wicked and air dried. EM grids were imaged at 120 kV on a FEI Tecnai Spirit TEM system. Images were acquired as 2048 × 2048 pixel, 16-bit gray scale files using the FEI’s TEM Imaging and Analysis (TIA) interface on an Eagle 2 K CCD multiscan camera. All the images were acquired at 2–4 microns underfocused to improve sample contrast.
6
Electron Microscopy of Extracellular Vesicles
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Electron microscopy was performed at the Multiscale Microscopy Core, an Oregon Health & Science University Shared Resource facility. Briefly, EV preparations were fixed in s 4% paraformaldehyde (PFA) solution (Thermo Scientific). Five microliters of fixed EVs were deposited onto glow discharged (60 s 15 mA, negative mode) carbon formvar 400 Mesh copper grids (Ted Pella 01822‐F) for 3 min, rinsed 30 s in water, wicked on Whatman filter paper 1, stained for 60 s in filtered 1.33% (w/v) uranyl acetate in water, wicked and air dried. Samples were imaged at 120 kV on a FEI Tecnai™ Spirit TEM system. Images were acquired using the AMT software interface on a NanoSprint™12S‐B CMOS camera system.
7
Isolation and Characterization of Extracellular Vesicles
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5 µl of SEC-isolated EVs were deposited onto glow discharged (120 s 15 mAmp, negative mode) carbon formvar 400 Mesh copper grids (Ted Pella 01822-F) for 3 min, rinsed for 15 s in water, wicked on Whatman filter paper 1, stained for 3 min in freshly prepared 1% (w/v) uranyl acetate in water, wicked and air dried. Samples were imaged at 120 kV on a FEI Tecnai™ Spirit TEM system. Images were acquired as 2048 × 2048 pixel, 16-bit gray scale files using the FEI’s TEM Imaging and Analysis (TIA) interface on an Eagle™ 2K CCD multiscan camera. Nanoparticle density assessed by flow cytometry in the EV fractions after size exclusion chromatography ranged from 2.4 to 3.6 × 105 events/µl.
8
Transmission Electron Microscopy Imaging of DNA
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Five microliters of DNA suspensions was deposited onto glow discharged (120 s 15 mAmp, negative mode) carbon formvar 400 Mesh copper grids (Ted Pella 01822-F) for 3 min, rinsed 10 s in water, wicked on Whatman filter paper 1, stained for 3 min in freshly prepared 1% (w/v) uranyl acetate in water, wicked, and air-dried. Samples were imaged at 120 kV on a FEI Tecnai Spirit TEM system. Images were acquired using the AMT interface on an AMT 12 Megapixel NanoSprint12S-B cMOS camera system.
9
Transmission Electron Microscopy of Nanoparticles
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Transmission
electron microscopy (TEM) was performed at 120 kV on a FEI Tecnai
Spirit TEM system. Images were acquired using the AMT software interface
on a NanoSprint12S-B cMOS camera system. TEM samples were typically
prepared by drop-casting 5 μL NP colloidal dispersions in water
onto 400 mesh copper TEM grids with carbon films (CF400-Cu, Electron
Microscopy Sciences). The samples were allowed to air-dry for 30 min
and kept overnight in a grid storage box (71150, Electron Microscopy
Sciences) until the imaging time.
electron microscopy (TEM) was performed at 120 kV on a FEI Tecnai
Spirit TEM system. Images were acquired using the AMT software interface
on a NanoSprint12S-B cMOS camera system. TEM samples were typically
prepared by drop-casting 5 μL NP colloidal dispersions in water
onto 400 mesh copper TEM grids with carbon films (CF400-Cu, Electron
Microscopy Sciences). The samples were allowed to air-dry for 30 min
and kept overnight in a grid storage box (71150, Electron Microscopy
Sciences) until the imaging time.
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