Fei tecnai spirit electron microscope

Manufactured by Thermo Fisher Scientific

The FEI Tecnai Spirit is a transmission electron microscope designed for high-resolution imaging and analysis of samples at the nanoscale. It is capable of providing detailed structural information about a wide range of materials, including biological specimens, thin films, and nanostructures. The Tecnai Spirit utilizes an electron beam to illuminate the sample and capture images, allowing users to study the morphology, composition, and crystalline structure of their samples with high magnification and resolution.

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Lab products found in correlation

Reichert ultracut s ultramicrotome, by Leica (2 mentions) Quemesa, by Thermo Fisher Scientific (2 mentions) Quemesa, by Olympus (1 mentions) Cacodylate buffer, by Electron Microscopy Sciences (1 mentions) Sodium cacodylate buffer, by Merck Group (1 mentions) Propylene oxide, by Merck Group (1 mentions) Osmium tetroxide, by Electron Microscopy Sciences (1 mentions) Em uc7 ultramicrotome, by Leica (1 mentions) Paraformaldehyde (pfa), by Electron Microscopy Sciences (1 mentions) Veleta, by Olympus (1 mentions) Ab9110, by Abcam (1 mentions) A11122, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

FEI Tecnai Spirit BioTWIN electron microscope Tecnai G2 Spirit 120 kV (FEI) electron microscope FEI Tecnai Spirit Biotwin transmission electron microscope FEI Tecnai G2 Spirit Twin 120 kV transmission electron microscope FEI Tecnai‐12 G2 Spirit Biotwin electron microscope FEI Tecnai G2 Spirit Twin Transmission electron microscope FEI Tecnai™ G2 Spirit BioTWIN transmission electron microscope Tecnai Spirit electron microscope Tecnai Spirit microscope Tecnai Spirit

6 protocols using fei tecnai spirit electron microscope

Ultrastructural Analysis of Melanocyte Organelles

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Control and Rab4A-knockdown melanocytes were seeded on Matrigel-coated glass coverslips. After 24 h, cells were fixed initially with 0.5% Karnovsky's fixative (4% paraformaldehyde, 72 mM sodium cacodylate pH 7.4, 4 mM CaCl₂, 0.5% glutaraldehyde) for 2 h followed by overnight fixation with 2% Karnovsky's fixative (contains 2% glutaraldehyde). Cells were processed for Epon embedding as described previously (Raposo et al., 2001 (link)). Ultrathin sections of cell monolayers were prepared with a Reichert UltracutS ultramicrotome (Leica Microsystems) and contrasted with uranyl acetate and lead citrate as described previously (Raposo et al., 2001 (link)). Samples were examined with a FEI Tecnai Spirit electron microscope (FEI Company), and digital acquisitions were made with a numeric camera (Quemesa; Soft Imaging System). For quantification, melanosome stages were defined by morphology (Raposo et al., 2001 (link)) and vacuoles were defined as empty organelles. The number of melanosomes and vacuoles per µm² cytosol were counted using ImageJ software. We counted ten cells from each control sh and Rab4A sh condition. Furthermore, we estimated the melanosome stages from 883 total melanosomes of control sh and 300 total melanosomes of Rab4A sh cells.

Nag S., Rani S., Mahanty S., Bissig C., Arora P., Azevedo C., Saiardi A., van der Sluijs P., Delevoye C., van Niel G., Raposo G, & Setty S.R. (2018). Rab4A organizes endosomal domains for sorting cargo to lysosome-related organelles. Journal of Cell Science, 131(18), jcs216226.

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Transmission Electron Microscopy of SKOV-3 Cells

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For TEM, 6 × 10⁵ WT or NFE2L2/NRF2 KO SKOV-3 cells were seeded in six-well plates and, 24 hours later, processed as described previously (65 (link)). Briefly, cells were fixed in 2.5% glutaraldehyde + 2% PFA in 0.1 M piperazine-N,N′-bis(2-ethanesulfonic acid) or Pipes buffer (pH 7.4) for 1 hour at room temperature. Samples were postfixed in 1% osmium tetroxide in Pipes buffer for 1 hour following a wash in 0.05 M Pipes buffer + 0.05 M glycine and 2 × 10 min washes in 0.1 M Pipes. Following two more 10-min washes in DIW, samples for TEM were block-stained in 2% aqueous uranyl acetate, washed in DIW, dehydrated through a graded series of alcohols, infiltrated with 1:1 alcohol and Spurr’s resin overnight, and then embedded in 100% Spurr’s resin overnight at 60°C. Sections were viewed using a FEI Tecnai Spirit electron microscope (FEI Company, Hillsboro, OR) operated at 100 kV. Eight-bit TIFF images were captured through an AMT 4-megapixel camera.

Anandhan A., Dodson M., Shakya A., Chen J., Liu P., Wei Y., Tan H., Wang Q., Jiang Z., Yang K., Garcia J.G., Chambers S.K., Chapman E., Ooi A., Yang-Hartwich Y., Stockwell B.R, & Zhang D.D. (2023). NRF2 controls iron homeostasis and ferroptosis through HERC2 and VAMP8. Science Advances, 9(5), eade9585.

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Ultrastructural Analysis of Melanocytes

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Control and Rab4A-knockdown melanocytes were seeded on Matrigel coated glass coverslips. Post 24 h, cells were fixed initially with 0.5% Karnovsky’s fixative (4% paraformaldehyde, 72 mM sodium cacodylate pH 7.4, 4 mM CaCl₂, 0.5% glutaraldehyde) for 2 h followed by overnight fixation with 2% Karnovsky’s fixative (contains 2% glutaraldehyde). Cells were processed for Epon embedding as described (Raposo et al., 2001 (link)). Ultrathin sections of cell monolayers were prepared with a Reichert UltracutS ultramicrotome (Leica Microsystems) and contrasted with uranyl acetate and lead citrate as described (Raposo et al., 2001 (link)). Samples were examined with a FEI Tecnai Spirit electron microscope (FEI Company), and digital acquisitions were made with a numeric camera (Quemesa; Soft Imaging System). For quantification, melanosome stages were defined by morphology (Raposo et al., 2001 (link)) and vacuoles were defined as empty organelles. Melanosomes and vacuoles per µm2 cytosol were counted using ImageJ software. We have counted 10 cells each from control sh and Rab4A sh condition. Further, we have estimated the melanosome stages from 883 total melanosomes of control sh and 300 total melanosomes of Rab4A sh cells.

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Ultrastructural Analysis of Brain Regions

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Brains were extracted and four regions, namely the midbrain, striatum, hippocampus, and cortex, were microdissected. Tissue slices for transmission (TEM) electron microscopy were fixed in 2.5% glutaraldehyde + 2% PFA in 0.1 M piperazine-N,N′-bis (2-ethanesulfonic acid) or PIPES buffer (pH 7.4) for 1 h at RT or at 4 °C overnight. Samples were post-fixed in 1% osmium tetroxide in PIPES buffer for 1 h following a wash in 0.05 M PIPES buffer +0.05 M Glycine and 2 × 10 min washes in 0.1 M PIPES. Following two more 10 min washes in deionized water (DIW), samples for TEM were block stained in 2% aqueous uranyl acetate, washed in DIW, dehydrated through a graded series of alcohols, infiltrated with 1:1 alcohol and Spurr’s resin overnight, and then embedded in 100% Spurr’s resin overnight at 60 °C. Sections were viewed using a FEI Tecnai Spirit electron microscope (FEI Company, Hillsboro, OR) operated at 100 kV. 8-bit TIFF images were captured through an AMT 4 Mpixel camera. Morphometric measurements were conducted in digital images using Image J (NIH) software. Assessment of mitochondrial morphology was performed as described previously [6 (link)].

Anandhan A., Chen W., Nguyen N., Madhavan L., Dodson M, & Zhang D.D. (2022). α-Syn overexpression, NRF2 suppression, and enhanced ferroptosis create a vicious cycle of neuronal loss in Parkinson’s disease. Free radical biology & medicine, 192, 130-140.

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Ultrastructural Analysis of Organoids

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Organoids were fixed in 2.5% glutaraldehyde (Electron Microscopy Sciences, #16220) and 2% PFA (Electron Microscopy Sciences, #15710) in 0.1 M sodium cacodylate buffer (Sigma, #20840) overnight at 4°C, washed with 0.1 M cacodylate buffer, and post-fixed in 1% osmium tetroxide (Electron Microscopy Sciences, #19170) for 2 hours at room temperature. Organoids were then dehydrated in a graded ethanol (Sigma, #51976) series (25%, 50%, 75%, and 100%), further dehydrated in propylene oxide (Sigma, #82320) and embedded in Epon resin (SERVA Electrophoresis; glycid ether #21045.01, dodecenylsuccinic acid anhydride #20755.01, methylnadic anhydride #29452.01 and benzyl dimethylamine #14835.01) for 12 h. Semi-thin (0.4 μm) and ultra-thin sections (50 nm) were cut with a Leica EM UC7 ultramicrotome and the latter were collected on formvar-coated single-slot copper grids (Electron Microscopy Sciences, #FF2010-Cu) for imaging. Sections were contrasted with 1% uranyl acetate (Electron Microscopy Sciences, #22400) and lead citrate (Electron Microscopy Sciences, #17800) (8 minutes each) and imaged on a FEI Tecnai Spirit electron microscope (FEI Company) operated at 120 kV using a side-mounted 2K × 2K CCD camera (Veleta, Olympus).

Cowan C.S., Renner M., De Gennaro M., Gross-Scherf B., Goldblum D., Hou Y., Munz M., Rodrigues T.M., Krol J., Szikra T., Cuttat R., Waldt A., Papasaikas P., Diggelmann R., Patino-Alvarez C.P., Galliker P., Spirig S.E., Pavlinic D., Gerber-Hollbach N., Schuierer S., Srdanovic A., Balogh M., Panero R., Kusnyerik A., Szabo A., Stadler M.B., Orgül S., Picelli S., Hasler P.W., Hierlemann A., Scholl H.P., Roma G., Nigsch F, & Roska B. (2020). Cell Types of the Human Retina and Its Organoids at Single-Cell Resolution. Cell, 182(6), 1623-1640.e34.

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Ultrastructural Analysis of HCMV-US28 and iHA-US28

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Doxycyclin-induced iHA-US28-U251 cells and HCMV-US28-pHluorin-infected HFFF-Tet cells were fixed in 2% PFA, 0.2% glutaraldehyde in 0.1M phosphate buffer pH 7.4. Cells were then washed with phosphate buffer, embedded in 10% (wt/vol) gelatin and infused in 2.3 M sucrose. Mounted gelatin blocks were frozen in liquid nitrogen and ultrathin sections were prepared with an Ultracut FCS ultracryomicrotome (Leica). Ultrathin cryosections were labeled with rabbit anti-HA (cat. #: ab9110, Abcam, 1:500) or rabbit anti-GFP (cat. #: A11122, Invitrogen, 1:200) antibodies, and protein A coupled to 10nm gold particles. EVs and virions were isolated from the culture medium of HCMV-infected HFFF-Tet cells. Culture medium was centrifuged at 500xg to remove dead cells and cell debris and subsequently centrifuged for 1 hour at 23,000 rpm to isolate EVs and virions. EVs and virions were spotted on carbon-coated and formvar coated EM grids and fixed with 2% PFA in PBS before staining with anti-GFP (antibody and protein A coupled to 10nm gold particles. Samples were examined with a FEI Tecnai Spirit electron microscope (FEI Company), and digital acquisitions were made with a numeric camera (Quemesa; Soft Imaging System).

Bebelman M.P., Setiawan I.M., Bergkamp N.D., van Senten J.R., Crudden C., Bebelman J.P., Verweij F.J., van Niel G., Siderius M., Pegtel D.M, & Smit M.J. (2023). Exosomal release of the virus-encoded chemokine receptor US28 contributes to chemokine scavenging. iScience, 26(8), 107412.

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