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Tecnai 12 twin tem

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The Tecnai 12 TWIN TEM is a transmission electron microscope (TEM) designed for high-resolution imaging and analysis of samples. It features a LaB6 electron source and a two-lens condenser system for improved illumination and resolution. The Tecnai 12 TWIN TEM is capable of operating in both TEM and STEM modes for a wide range of applications in materials science, biology, and nanotechnology.

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Lab products found in correlation

Synergy 2, by Agilent Technologies (1 mentions) Malvern zetasizer, by Malvern Panalytical (1 mentions) Vitrobot, by Thermo Fisher Scientific (1 mentions) Lc325 cu, by Electron Microscopy Sciences (1 mentions) Cell observer system, by Zeiss (1 mentions) Microtome, by Leica (1 mentions) Tannic acid, by Thermo Fisher Scientific (1 mentions) Diamond knife, by Electron Microscopy Sciences (1 mentions) Odyssey imaging system, by LI COR (1 mentions) Oneview camera model 1095, by Ametek (1 mentions)

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Tecnai 12 (254 citations) Tecnai 12 TEM (44 citations) Tecnai 12 G2 TWIN TEM (6 citations) Tecnai 12 Spirit Bio TWIN TEM (3 citations) Tecnai G2 12 Twin TEM 120 kV (3 citations) Tecnai G2 Spirit Twin T-12 TEM (2 citations)

10 protocols using tecnai 12 twin tem

1

Characterization of RBC Membrane-Coated Nanoparticles

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Nanoparticles coated with RBC membranes were characterized by TEM using a FEI Tecnai 12 TWIN TEM. The membrane-coated nanoparticles were stained in 1% uranyl formate as a negative stain. The particles were also characterized by DLS using a Malvern ZetaSizer (Malvern Instruments, Westborough, MA). The particles were suspended at a concentration of 1 mg/ml in 1× PBS and sized in a low-volume disposable cuvette using recommended machine settings. Zeta potential of the particles was measured using a Malvern ZetaSizer Pro. The particles were suspended at a concentration of 1 mg/ml in deionized water and sized in a disposable folded capillary cell using recommended machine settings. To evaluate the presence of CD47 on the surface of the particle, allophycocyanin-labeled anti-mouse CD47 antibody was used (BioLegend, San Diego, CA). Coated particles and uncoated spherical particles were incubated with a 1:100 dilution of the antibody for 1 hour at 4°C in 1× PBS. Following incubation, the particles were washed three times in 1× PBS at 17,000g for 5 min and read on a BioTek Synergy 2 plate reader (BioTek, Winooski, VT).
Ben-Akiva E., Meyer R.A., Yu H., Smith J.T., Pardoll D.M, & Green J.J. (2020). Biomimetic anisotropic polymeric nanoparticles coated with red blood cell membranes for enhanced circulation and toxin removal. Science Advances, 6(16), eaay9035.
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2

CryoTEM Analysis of Protein Morphology

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CryoTEM was performed to determine morphology in the presence of aqueous buffer. Briefly, cryoTEM specimens were prepared using an FEI Vitrobot (Hillsboro, OR). Protein solutions were kept in an ice bath (4 °C) before processing and then raised to 37 °C immediately prior to blotting. Six microliters of sample were pipetted onto a TEM grid coated with a lacey carbon film (LC325-Cu, Electron Microscopy Sciences). The specimen was then blotted under 95% humidity, immediately transferred into liquid ethane, and stored in liquid nitrogen environment. Micrographs were acquired using FEI Tecnai 12 TWIN TEM equipped with 16 bit 2Kx2K FEI eagle bottom mount camera (Hillsboro, OR). All cryoTEM images were acquired at an accelerating voltage of 100 kV. Images were analyzed using ImageJ (NIH, USA).
Aluri S.R., Shi P., Gustafson J.A., Wang W., Lin Y.A., Cui H., Liu S., Conti P.S., Li Z., Hu P., Epstein A.L, & MacKay J.A. (2014). A Hybrid Protein–Polymer Nanoworm Potentiates Apoptosis Better than a Monoclonal Antibody. ACS Nano, 8(3), 2064-2076.
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3

Visualizing Cellular Uptake of SPIO Nanoparticles

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SPIO labeled cells were stained for the presence of internalized iron oxide particles using a Prussian Blue stain (10 (link)). Using this method, the magnetic particles are converted to an intense dark blue color following 30 min incubation with Perls' reagent (1 gram potassium ferrocyanide in 42 ml deionized water + 8 ml 37.5 % HCl). SPIO uptake in washed, labeled 4% glutaraldehyde-fixed cells was visualized using a Zeiss Cell Observer System with Apotome 2 Digital Imaging. For transmission electron microscopy TEM), washed, labeled cell samples were fixed with 3.0% formaldehyde, 1.5% glutaraldehyde in 0.1M sodium cacodylate, 5 mM Ca2+, and 2.5% sucrose. Following staining with Palade's OsO4 and uranyl acetate, cellular uptake in Epon-embedded cell samples was analyzed with a FEI Tecnai 12 TWIN TEM operating at 100kV.
Bulte J.W., Walczak P., Janowski M., Krishnan K.M., Arami H., Halkola A., Gleich B, & Rahmer J. (2015). Quantitative “Hot-Spot” Imaging of Transplanted Stem Cells Using Superparamagnetic Tracers and Magnetic Particle Imaging. Tomography, 1(2), 91-97.
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4

LTBP-1 Interaction with Fibrillin Microfibrils

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Full-length human LTBP-1 was purified as previously described56 (link) and incubated with bovine fibrillin microfibrils at a 2:1 molar ratio for 4 h at 4 °C. The complex was adhered for 60 s to 400-mesh carbon-coated grids then washed with Milli-Q water three times and dried. The sample was visualized in STEM mode with a Fischione high-angle annular dark-field detector on an FEI Tecnai 12 Twin TEM at 34,000× magnification. A camera length of 350 cm was used to give an angular collection range of 20–100 mrad. Tobacco mosaic virus was used as a calibration standard for mass per unit length. The electron dose was kept sufficiently low (<300 e nm−1) to produce negligible mass loss. Mass per unit length measurements and axial mass distributions were measured from STEM annular dark-field images using the Semper6 image analysis software (Synoptics).
Godwin A.R., Dajani R., Zhang X., Thomson J., Holmes D.F., Adamo C.S., Sengle G., Sherratt M.J., Roseman A.M, & Baldock C. (2023). Fibrillin microfibril structure identifies long-range effects of inherited pathogenic mutations affecting a key regulatory latent TGFβ-binding site. Nature Structural & Molecular Biology, 30(5), 608-618.
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5

Visualizing hBMEC Monolayer Barrier Properties

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TEM was used to characterize the monolayer of hBMECs in the in vitro BBB model with and without exposure to nanoparticles in order to visualize the barrier properties, i.e. validation of tight junctions (TJs) and to evaluate mechanisms of nanoparticle crossing. As preparation of cross-sections for TEM characterization, the transwells containing a monolayer of hBMECs were first fixed with 3.0% formaldehyde and 1.5% glutaraldehyde in a 0.1 M cacodylate buffer, pH 7.4, at room temperature for one hour. Washing in 0.1 M cacodylate buffer was carried out for 15 min two times prior to post-fixation in Palade’s 1% osmium tetroxide (OsO4) for one hour on ice. A washing in Milli-Q H2O was then applied for 15 min followed by further staining for 30 min using 1% tannic acid (Fisher Scientific) in 0.1 M cacodylate buffer. The samples were thereafter incubated overnight in Kellenberger solution, and subsequently rinsed once in Milli-Q H2O and once in 50% ethanol. Dehydration was performed in graded series of ethanol (70%, 95%, and 100%), and then three washes in 100% ethanol prior to embedding in Epon. A diamond knife (Diatome, Hatfield, PA, USA) on a Leica Microtome (Leica Biosystems, Buffalo, IL, USA) was used to cut ultrathin (80 nm) sections. TEM imaging was carried out using a FEI Tecnai 12 TWIN TEM (FEI) microscope operated at 120 kV.
Karlsson J., Rui Y., Kozielski K.L., Placone A.L., Choi O., Tzeng S.Y., Kim J., Keyes J.J., Bogorad M.I., Gabrielson K., Guerrero-Cazares H., Quiñones-Hinojosa A., Searson P.C, & Green J.J. (2019). Engineered nanoparticles for systemic siRNA delivery to malignant brain tumours. Nanoscale, 11(42), 20045-20057.
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6

Negative Staining of Amyloid Fibrils

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5 μL aliquots from samples used in the ThT experiments (after 34 h incubation) were placed on 400-mesh copper grids covered with a carbon-stabilized Formvar film. Excess solutions were removed following 2 min of incubation, and the grids were negatively stained for 30 s with a 1% uranyl acetate solution. Samples were viewed in an FEI Tecnai 12 TWIN TEM operating at 120 kV.
Bloch D.N., Sandre M., Ben Zichri S., Masato A., Kolusheva S., Bubacco L, & Jelinek R. (2023). Scavenging neurotoxic aldehydes using lysine carbon dots. Nanoscale Advances, 5(5), 1356-1367.
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7

Characterizing Conjugate Size and Efficiency

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Dye conjugation efficiency and stoichiometry was confirmed by absorbance at 280/745 nm. Conjugate sizes were compared by 10% PAGE (BioRad Lab Inc., Hercules, CA; 200 ng of each J591-conjugates), visualized by LICOR Odyssey imaging system (LI-COR Biosciences, Lincoln, NE). Sizes were estimated by primary amines and 75% conjugation efficiency. Sizes of PEGylated conjugates were determined by Transmission Electron Microscopy (TEM). Bright-field images were taken on an FEI Tecnai 12 TWIN TEM equipped with 16 bit 2K x2K FEI Eagle bottom mount camera and SIS Megaview III wide-angle camera. TEM grids were prepared by drying 10 μL of sample on copper grids with holey carbon films overnight (grids from Ted Pella). Particle size was determined using ImageJ, where at least 23 particles were analyzed per sample.
Mukherjee A., Kumar B., Hatano K., Russell L.M., Trock B.J., Searson P.C., Meeker A.K., Pomper M.G, & Lupold S.E. (2016). Development and application of a novel model system to study ‘active’ and ‘passive’ tumor targeting. Molecular cancer therapeutics, 15(10), 2541-2550.
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8

Characterization of HBc VLP Vaccine

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To analyze the average size and zeta potential, the HBc VLP vaccine solution was measured using a Malvern Zetasizer (Nano ZS, Malvern, UK). The morphology of our HBc VLP vaccine was investigated by transmission electron microscopy (TEM). Images of samples were obtained using a Tecnai 12 Twin TEM (FEI Company, Hillsboro, OR, USA) equipped with a OneView Camera Model 1095 (Gatan, Pleasanton, CA, USA) and a voltage of 120 kV.
Hao Y., Gu Z., Yu Z., Schomann T., Sayedipour S., Aguilar J.C., ten Dijke P, & Cruz L.J. (2022). Photodynamic Therapy in Combination with the Hepatitis B Core Virus-like Particles (HBc VLPs) to Prime Anticancer Immunity for Colorectal Cancer Treatment. Cancers, 14(11), 2724.
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9

Characterization of Nanostructure Morphology

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FF nanostructures which were formed at either 1 mg/ml (above their critical concentration) or 0.5 mg/ml (under their critical concentration) were both diluted to 0.125 mg/ml and 10 µl of each sample was immediately subjected to high-resolution transmitting electron microscopy utilizing a Tecnai 12 TWIN TEM (FEI) transmitting electron microscope. Micrographs were obtained for three separate repetitions of each dilution and 50 nanostructures were measured for both their width and length (a total of 150 nanostructures were measured). The results of this analysis can be found in Supplementary Fig. 2.
Schnaider L., Brahmachari S., Schmidt N.W., Mensa B., Shaham-Niv S., Bychenko D., Adler-Abramovich L., Shimon L.J., Kolusheva S., DeGrado W.F, & Gazit E. (2017). Self-assembling dipeptide antibacterial nanostructures with membrane disrupting activity. Nature Communications, 8, 1365.
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10

Negatively Stained TEM Imaging

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5 μL aliquots of samples used in the ThT experiments (after 20 h incubation) were placed on 400-mesh copper grids coated with a carbon-stabilized formvar film. Excess solutions were removed following 2 min incubation, and the grids were negatively stained for 30 s with a 1% uranyl acetate solution. Samples were viewed in a FEI Tecnai 12 TWIN TEM operating at 120 kV.
Bloch D.N., Ben Zichri S., Kolusheva S, & Jelinek R. (2020). Tyrosine carbon dots inhibit fibrillation and toxicity of the human islet amyloid polypeptide. Nanoscale Advances, 2(12), 5866-5873.
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