KSHV virion (concentration was 2 × 108/ml, 20 μl) was mixed with or without nAg at final concentration of 0.2 μg/ml and maintained at 37 °C for 2 h. A copper mesh with carbon support film was placed over the sample droplets (50 μl sample containing 107/ml of virus particles) and allowed to float for 3–10 min to ensure that the virus particles could adsorb onto the support film. The copper mesh was then removed from the sample droplets and placed on filter paper for liquid absorption, followed by staining on the 2% phosphotungstic acid-dyed droplets, and floating for 3 min. After absorbing the liquid, the copper mesh was dried under an incandescent lamp for 10 min and then observed under a FEI Tecnai G2 Spirit transmission electron microscopy (Thermo Fisher Scientific, USA).
Fei tecnai g2 spirit transmission
Manufactured by Thermo Fisher Scientific
Sourced in United States, Germany
The FEI Tecnai G2 Spirit transmission electron microscope is a high-performance instrument designed for advanced imaging and analysis of samples at the nanoscale level. It features a LaB6 electron source, providing high-resolution imaging capabilities, and supports a range of advanced analytical techniques.
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Lab products found in correlation
Eagle 4k hr 200kv, by Thermo Fisher Scientific (2 mentions)
P35g 1.5 14 cgrd, by Thermo Fisher Scientific (2 mentions)
Uct ultramicrotome, by Leica (2 mentions)
A1r laser scanning confocal microscope, by Nikon (2 mentions)
Lysotracker red, by Thermo Fisher Scientific (2 mentions)
Uranyl acetate, by Merck Group (1 mentions)
4 protocols using fei tecnai g2 spirit transmission
1
Visualizing KSHV Virion Interactions
2
Histological Analysis of Testicular Tissues
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Gonads were fixed in 4% paraformaldehyde solution, embedded in paraffin (Shenggong Co, Ltd), and cross sections were cut at 5-μm thickness for H&E staining as previously described (20 ). Small blocks of various testes were fixed in a mixture of 2.5% (v/v) glutaraldehyde in PBS (4 °C, pH 7.4, 0.1 M), overnight, postfixed in 1% (w/v) osmium tetroxide PBS for 1 h at RT, dehydrated in a graded ethanol series, and embedded in Epon 812. Ultrathin sections of 70-nm thickness were prepared using diamond knives on Leica EM UC7 Ultramicrotome, stained with 2% uranyl acetate (Merck, Darmstadt, Germany) and 2.8% lead citrate for 10 min per step, and examined with a FEI Tecnai G2 Spirit transmission electron microscope (Thermo Fisher) at an accelerating voltage of 30 kV.
3
Correlative Light-Electron Microscopy of Lysosomes
4
Correlative Light-Electron Microscopy of Lysosomes
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For electron microscopy (EM), cells were grown on coverslips and fixed in a mixture of 2.5% glutaraldehyde and 2% paraformaldehyde in 0.1M cacodylate buffer for 2–24 h at 4 °C. After post-fixation in 1% osmium tetroxide and 3% uranyl acetate, cells were dehydrated in series of ethanol, embedded in Epon resin and polymerized for 48 h at 60 °C. Ultrathin sections were made using UCT ultramicrotome (Leica Microsystems) and contrasted with 4% uranyl acetate and Reynolds’s lead citrate. Samples were imaged using a FEI Tecnai Spirit G2 transmission electron microscope (FEI Company, Hillsboro, OR) operated at 80 kV. Images were captured with an Eagle 4k HR 200kV CCD camera. For correlative light electron microscopy, cells were grown on gridded glass bottom culture dishes (MatTek; P35G-1.5-14-CGRD) and incubated for 45 min with LysoTracker Red (2 μM) (Thermo Fisher) prior to EM fixation. Fixed cells were imaged on the Nikon A1R laser scanning confocal microscope for LysoTracker staining using z-stacks with step sizes of 0.2 μm as described above, and subsequently processed and imaged for EM as described above.
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