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Tecnai g2 tf20 super twin

Manufactured by Thermo Fisher Scientific

The Tecnai G2 TF20 Super TWIN is a high-performance transmission electron microscope (TEM) designed for advanced materials analysis. It features a super-twin lens configuration for enhanced resolution and imaging capabilities. The core function of this product is to enable detailed structural and compositional analysis of a wide range of materials at the nanoscale level.

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7 protocols using tecnai g2 tf20 super twin

1

Transmission Electron Microscopy of Samples

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Samples were fixed with 1% (w/v) glutaraldehyde in PBS for 10 min, then loaded on a carbon/formvar-coated grid (5 µl each sample) and dried under vacuum for 20 min. Samples were incubated with 2% uranyl acetate (10 µl) for 30 s, then dried under vacuum for 20 min. The ultrastructure of each sample was observed by a transmission electron microscopy (FEG-TEM, FEI Tecnai G2 TF20 Super TWIN).
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2

Microscopy Techniques for Cell Synchronization

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For immunofluorescence confocal microscopy, three-dimensional structured illumination microscopy (3D-SIM), and electron microscopy, cells were processed as previously described32 (link). Briefly, cells on coverslips were treated with aphidicolin for synchronization at the G1/S phase transition and released with fresh medium at the indicated times. The cells were fixed in methanol and incubated with the indicated primary antibodies. After wash, the cells were incubated with Alexa Fluor 488-, Alexa Fluor 568-, or Alexa Fluor 647-conjugated secondary antibodies (Invitrogen). In some experiments, cells were immunostained with the Alexa Fluor 568-conjugated CEP120 antibody. The samples were viewed on a confocal system (LSM 700 or LSM880 Airyscan systems; Carl Zeiss). The 3D-SIM super-resolution images were acquired using a Zeiss ELYRA system equipped with a Plan Apochromat 63 × /1.4NA oil-immersion objective and the ZEN software (Carl Zeiss). For electron microscopy (EM), cells were grown on Aclar film (Electron Microscopy Sciences), fixed, and embedded in Spurr’s resin as previously described32 (link). Thin sections (100 nm) were stained with 4% uranyl acetate and Reynold’s lead citrate and the samples were examined with an electron microscope (T FEG-TEM; FEI Tecnai G2 TF20 Super TWIN)32 (link).
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3

Immunogold Labeling of ATP5A

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Thin sections placed on nickel grids were blocked with 5% BSA in PBS for 20 min and incubated with the mouse anti-ATP5A (500x, ab14748, Abcam) antibodies in incubation buffer (1% BSA in PBS) for 2 h. Grids were subsequently washed with incubation buffer three times (10 min each). Secondary antibodies, goat anti-mouse IgG (EM.GMHL15, BB International) with 15 nm gold conjugations at 20-fold dilution were further applied and incubated for 1 h. After washing with PBS, the immune-complexes were fixed with 1% glutaraldehyde in PBS for 5 min and washed three times with distilled water (10 min each). The specimens were inspected by TEM operating at 120 kV (FEI Tecnai G2 TF20 Super TWIN).
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4

Immunogold Labeling of Cellular Organelles

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Thin sections placed on nickel grids were blocked with 10% BSA in PBS for 20 min, and incubated with primary antibodies in incubation buffer (1% BSA in PBS) for 2 hr. Grids were subsequently washed with incubation buffer three times (10 min each). Secondary antibodies, goat anti-mouse IgG (EM.GMHL15, BB International) and protein A (EM.PAG15, BB International) conjugated to 15 nm gold particles, were used against the primary antibodies from mouse and rabbit respectively. Secondary antibodies at 20-fold dilution were applied and samples were incubated for 1 hr. After washing with PBS, the immune-complexes were fixed with 1% glutaraldehyde in PBS and washed three times with distilled water. The specimens were inspected by TEM operating at 120 kV (FEI Tecnai G2 TF20 Super TWIN).
The primary antibodies and applied dilution factors are listed as follows: mouse anti-dsDNA (500x, abcam ab27156), mouse anti-ATP5A (500x, abcam ab14748), mouse anti-Cytochrome C (8000x, abcam ab13575), mouse anti-PDHA1 (500x, abcam ab110334), mouse anti-Ubiquitin (1000x, abcam ab7254), mouse anti-DNA-RNA hybrid (500x, kerafast ENH001), and rabbit anti-SOD2 (500x, abcam ab13534).
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5

Visualization of White Spot Syndrome Virus

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The original WSSV strain was collected from WSSV-infected Penaeus monodon in Taiwan [11 (link)]. The virus was then proliferated and purified in Procambarus clarkii as described in previous studies [6 (link)]. To visualize the virus particles, negative staining was performed on the 150-mesh grids with carbon films. Glow discharge was applied to the film to make a hydrophilic surface. Then, 10 μL of the virus solution was dropped onto the carbon films for 1 min absorption. After blotting out the excess solution, the grids were quickly rinsed and blotted through 3 drops of distilled water and stained with 0.35% uranyl formate for 1 min. The grids were allowed to be air-dried after the blotting of staining solutions. Images were taken under the transmission electron microscopy (TEM) operating at 120 kV (FEI Tecnai G2 TF20 Super TWIN).
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6

Transmission Electron Microscopy Imaging

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The samples were dropped onto 400-mesh Formvar carbon-coated copper grids (EMS Electron Microscopy Sciences, Hatfield, PA, USA) and incubated for 5 mins and washed twice by double distilled water, then negatively staining was performed with 2% uranyl acetate for 2 mins and dried at room temperature overnight. Image were observed with a FEG-TEM, FEI Tecnai G2 TF20 Super TWIN transmission electron microscope with an accelerating voltage of 120 kV.
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7

Transmission Electron Microscopy of FTH1

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FTH1 samples were prepared in 1X PBS (pH 8.0) to final concentrations of 0.075 mg/mL 5 μL of samples were dropped onto TEM grids (formvar/carbon 400 mesh, copper) followed by glow discharge treatments (Emitech K100X, 25 mA for 30 s). After air drying for 90 s, excess solution was wicked with filter paper. Next, the grids were rinsed with 5 µL of distilled deionized water followed by wicking with filter paper. This step was repeated in triplicate. 5 μL filtered 1% uranyl acetate was dropped onto grids, incubated for 60 s, then wicked with filter paper. The grids were kept in a grid-box and stored in an electronic dry cabinet until the grids were fully dried. The TEM images of FTH1 variants were taken by the FEI Tecnai G2 TF20 Super-Twin. The accelerating voltage was set as 120 kV.
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