Tecnai g2 spirit

Manufactured by Ametek

The Tecnai G2 Spirit is a transmission electron microscope (TEM) designed for high-resolution imaging and analysis of a wide range of materials. It features a LaB6 electron source, advanced optical systems, and a modern digital imaging platform to provide exceptional performance and image quality. The Tecnai G2 Spirit is a versatile instrument suitable for various applications in materials science, life sciences, and nanotechnology research.

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Lab products found in correlation

Ultrascan 4000 4k 4k ccd camera, by Ametek (3 mentions) Ultrascan 4000 4k x 4k ccd camera, by Ametek (3 mentions) Ultrascan 4000 ccd camera, by Ametek (3 mentions) Ultrascan 4000 4k 4k, by Ametek (2 mentions) Filter paper, by Cytiva (1 mentions) Whatman, by Cytiva (1 mentions) Methylcellulose, by Merck Group (1 mentions) Ultrascan 4000 ccd, by Ametek (1 mentions) Orius ccd camera, by Ametek (1 mentions)

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Tecnai G2 Spirit BioTwin (11 citations) Tecnai G2 Spirit BioTWIN TEM (5 citations) Tecnai G2 Spirit microscope (4 citations) Tecnai G2 Spirit electron microscope (4 citations) Tecnai Spirit (2 citations) Tecnai G2 Spirit TWIN Transmission Electron Microscope (2 citations) Tecnai G2 Spirit 120 kV TEM (2 citations) Tecnai G2 Spirit BioTWIN transmission electron microscope (2 citations) Tecnai G2 Spirit TEM (2 citations)

15 protocols using tecnai g2 spirit

Cryo-EM Imaging of Umb1 Particles

Cited 1 time

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Purified Umb1 particles were diluted to 0.01 mg ml^–1 and immediately subject to adsorption to glow-discharged carbon-coated copper grids for 60 s followed by 2% uranyl formate (w/v) staining. Micrographs were recorded using Leginon^{43 (link)} on a 120 KV FEI Tecnai G2 Spirit with a Gatan Ultrascan 4,000 4k × 4k CCD camera at ×67,000 nominal magnification. The defocus ranged from −1.0 to −2.0 µm and the pixel size was 1.6 Å. The parameters of the contrast transfer function (CTF) were estimated using CTFFIND^{44 (link)}. All particles were picked in a reference-free manner using DoG Picker^{45 (link)}. The particle stack from the micrographs was pre-processed in Relion⁴⁶. Particles were re-extracted with a binning factor of 4, resulting in a final box size of 80 pixels and a final pixel size of 6.4 Å. The reference-free 2D classification was performed using CryoSPARC^{47 (link)}.

Zhao Q., Bertolli S., Park Y.J., Tan Y., Cutler K.J., Srinivas P., Asfahl K.L., Fonesca-García C., Gallagher L.A., Li Y., Wang Y., Coleman-Derr D., DiMaio F., Zhang D., Peterson S.B., Veesler D, & Mougous J.D. (2024). Streptomyces umbrella toxin particles block hyphal growth of competing species. Nature, 629(8010), 165-173.

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Nanoparticle Imaging via Uranyl Formate Staining

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Nanoparticles diluted to 0.01 mg/mL in 50 mM Tris pH 8, 150 mM NaCl, were adsorbed to glow-discharged home-made carbon-coated copper grids for 30 seconds. The excess liquid was blotted away with filter paper (Whatman 1) and 3 μL of 2% w/v uranyl formate stain were applied to the grids. Finally, the stain was blotted away, and the grids were allowed to air dry for 1 min. Grids were imaged on a 120kV FEI Tecnai G2 Spirit with a Gatan Ultrascan 4000 4k x 4k CCD camera at 67,000 nominal magnification using a defocus ranging between 1.0 and 2.0 μm and a pixel size of 1.6 Å.

Tortorici M.A., Walls A.C., Joshi A., Park Y.J., Eguia R.T., Miranda M.C., Kepl E., Dosey A., Stevens-Ayers T., Boeckh M.J., Telenti A., Lanzavecchia A., King N.P., Corti D., Bloom J.D, & Veesler D. (2022). Structure, receptor recognition, and antigenicity of the human coronavirus CCoV-HuPn-2018 spike glycoprotein. Cell, 185(13), 2279-2291.e17.

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Negative Staining of Protein Samples

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Protein samples were adsorbed to glow-discharged carbon-coated copper grids for 30 s prior to 2% uranyl formate staining. Micrographs were recorded using the Leginon software^{71 (link)} on a 120 kV FEI Tecnai G2 Spirit with a Gatan Ultrascan 4000 CCD camera at 67,000 nominal magnification. The defocus ranged from 1.0 to 2.0 µm, and the pixel size was 1.6 Å.

Tortorici M.A., Walls A.C., Lang Y., Wang C., Li Z., Koerhuis D., Boons G.J., Bosch B.J., Rey F.A., de Groot R.J, & Veesler D. (2019). Structural basis for human coronavirus attachment to sialic acid receptors. Nature Structural & Molecular Biology, 26(6), 481-489.

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Negative Stain Transmission Electron Microscopy

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Protein samples were diluted to 0.01 mg/mL immediately prior to adsorption to glow-discharged carbon-coated copper grids for ~30 sec prior to a 2% uranyl formate staining. Micrographs were recorded using the Leginon software on a 120 kV FEI Tecnai G2 Spirit with a Gatan Ultrascan 4000 4 k x 4 k CCD camera at 67,000 nominal magnification. The defocus ranged from −1.0 to −2.0 μm and the pixel size was 1.6 Å.

Bowen J.E., Park Y.J., Stewart C., Brown J.T., Sharkey W.K., Walls A.C., Joshi A., Sprouse K.R., McCallum M., Tortorici M.A., Franko N.M., Logue J.K., Mazzitelli I.G., Nguyen A.W., Silva R.P., Huang Y., Low J.S., Jerak J., Tiles S.W., Ahmed K., Shariq A., Dan J.M., Zhang Z., Weiskopf D., Sette A., Snell G., Posavad C.M., Iqbal N.T., Geffner J., Bandera A., Gori A., Sallusto F., Maynard J.A., Crotty S., Van Voorhis W.C., Simmerling C., Grifantini R., Chu H.Y., Corti D, & Veesler D. (2022). SARS-CoV-2 spike conformation determines plasma neutralizing activity elicited by a wide panel of human vaccines. Science Immunology.

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Cryo-EM Protein Sample Preparation

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Protein was adsorbed to glow-discharged carbon-coated copper grids for 1 min prior to a 3X wash with water and 2% uranyl formate staining. Micrographs were recorded using the Leginon software(Suloway et al., 2005 (link)) on a 100kV FEI Tecnai G2 Spirit with a Gatan Ultrascan 4000 4k x 4k CCD camera at 67,000 nominal magnification. The defocus ranged from 1.0 to 2.0 μm and the pixel size was 1.6 Å. Particles were picked automatically in a reference free manner using DogPicker. Contrast transfer function (CTF) estimation was performed using GCTF(Zhang, 2016 (link)). Class averages were generated using Relion2.1(Kimanius et al., 2016 ).

Walls A.C., Xiong X., Park Y.J., Tortorici M.A., Snijder J., Quispe J., Cameroni E., Gopal R., Dai M., Lanzavecchia A., Zambon M., Rey F.A., Corti D, & Veesler D. (2019). Unexpected Receptor Functional Mimicry Elucidates Activation of Coronavirus Fusion. Cell, 176(5), 1026-1039.e15.

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Characterizing Extracellular Vesicle Morphology

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EV presence and morphology were characterised using transmission electron microscopy (TEM). 10 μL of each sample was placed onto a carbon coated glow discharged grid and incubated at room temperature for 20 min. Samples were then subject to a negative staining protocol. EVs were fixed onto the grid with 1% glutaraldehyde for 5 min. The sample grids were incubated on 1% aqueous uranyl acetate (UA) (Thermofisher Scientific, Massachusetts, USA), for 60 s, followed by 4% UA/2% methyl cellulose (Sigma Aldrich, Gillingham, UK) at a 1 : 9 ratio on ice for 10 min. Grids were then removed with a 5 mm wire loop and dried. The prepared grids were then viewed at 120 KV on a FEI Tecnai G2 Spirit with Gatan RIO16 digital camera.

Clarke E.J., Lima C., Anderson J.R., Castanheira C., Beckett A., James V., Hyett J., Goodacre R, & Peffers M.J. (2022). Optical photothermal infrared spectroscopy can differentiate equine osteoarthritic plasma extracellular vesicles from healthy controls. Analytical Methods, 14(37), 3661-3670.

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Negative Staining and Electron Microscopy Imaging

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Protein preparations were applied to glow-discharged continuous carbon EM grids and negatively stained with 2% uranyl formate. Grids were imaged by transmission electron microscopy using an FEI Tecnai G2 Spirit at 120kV acceleration voltage and a Gatan Ultrascan 4000 CCD using the Leginon software package (Suloway et al., 2009 (link)). Micrographs were collected at a nominal 67,000x magnification (pixel size 1.6 Å). GCTF was used for contrast transfer function (CTF) estimation, and Relion for particle picking and 2D classification (Scheres, 2012 (link); Zhang, 2016 (link); Zivanov et al., 2018 (link)).

Johnson M.C, & Kollman J.M. (2020). Cryo-EM structures demonstrate human IMPDH2 filament assembly tunes allosteric regulation. eLife, 9, e53243.

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Cryo-EM Protocol for Protein Imaging

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Protein was adsorbed to glow-discharged carbon-coated copper grids for 1 min prior to a 3X wash with water and 2% uranyl formate staining. Micrographs were recorded using the Leginon software (Suloway et al., 2005 (link)) on a 100kV FEI Tecnai G2 Spirit with a Gatan Ultrascan 4000 4k x 4k CCD camera at 67,000 nominal magnification. The defocus ranged from 1.0 to 2.0 μm and the pixel size was 1.6 Å. Particles were picked automatically in a reference free manner using DogPicker. Contrast transfer function (CTF) estimation was performed using GCTF (Zhang, 2016 (link)). Class averages were generated using Relion2.1 (Kimanius et al., 2016 ).

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Negative Staining for TEM Imaging

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Protein samples were adsorbed to glow-discharged carbon-coated copper grids for 30 seconds prior to 2% uranyl formate staining. Micrographs were recorded using the Leginon software^{72 (link)} on a 120kV FEI Tecnai G2 Spirit with a Gatan Ultrascan 4000 CCD camera at 67,000 nominal magnification. The defocus ranged from 1.0 to 2.0 μm and the pixel size was 1.6 Å.

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Negative Staining Electron Microscopy of E. coli and F. novicida Proteins

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Protein samples purified from E. coli were diluted to a concentration of 0.04 mg/mL and then adsorbed to glow-discharged carbon-coated copper mesh grids. Proteins isolated from F. novicida were purified as described in the immunoprecipitation section above, eluted with 100 μg/mL VSVG peptide and directly adsorbed without further dilution. Samples were incubated on the grids for 30 seconds prior to staining with 2% uranyl formate. Micrographs were acquired using the Leginon software (Suloway et al., 2005 (link)) on a 100kV FEI Tecnai G2 Spirit with a Gatan Ultrascan 4000 4k × 4k CCD camera at 52,000 nominal magnification (pixel size of 2.07 Å at the specimen level). The defocus ranged from 1.0 to 2.0 μm. The details of image analysis can be found in Supplemental Experimental Procedures.

Eshraghi A., Kim J., Walls A.C., Ledvina H.E., Miller C., Ramsey K.M., Whitney J.C., Radey M.C., Peterson S.B., Ruhland B.R., Tran B.Q., Goo Y.A., Goodlett D.R., Dove S.L., Celli J., Veesler D, & Mougous J.D. (2016). Secreted Effectors Encoded Within and Outside of the Francisella Pathogenicity Island Promote Intramacrophage Growth. Cell host & microbe, 20(5), 573-583.

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