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Transmission electron microscopy

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Transmission electron microscopy (TEM) is a powerful analytical technique that uses a focused beam of high-energy electrons to produce detailed images and analyze the structure and composition of materials at the nanoscale. The core function of TEM is to transmit an electron beam through a thin specimen, where the interaction between the electrons and the sample provides information about the sample's internal structure, chemical composition, and physical properties.

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Lab products found in correlation

Glutaraldehyde, by Merck Group (2 mentions) Thermomixer, by Eppendorf (1 mentions) Epon 812 resin, by Merck Group (1 mentions) Zetasizer nano, by Malvern Panalytical (1 mentions) Ems 200 cu, by Nisshin EM (1 mentions) Qwin image analysis software package, by Leica camera (1 mentions) Jsm 840 microscope, by JEOL (1 mentions) Gel electrophoresis, by Bio-Rad (1 mentions) Mass spectrometry, by Bruker (1 mentions) Inductively coupled plasma optical emission spectrometry, by Thermo Fisher Scientific (1 mentions) Multiskan spectrum microplate spectrophotometer, by Thermo Fisher Scientific (1 mentions) Zetaview, by Particle Metrix (1 mentions) Bca protein assay kit, by Beyotime (1 mentions) Dmem f12, by BD (1 mentions) Megaview 3 ccd camera, by Olympus (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Scanning electron microscopy, by Zeiss (1 mentions) Zetasizer 3000 ha, by Malvern Panalytical (1 mentions) Maldi tof mass spectrometer, by Shimadzu (1 mentions) Embed 812 epoxy, by Electron Microscopy Sciences (1 mentions)

52 protocols using transmission electron microscopy

1

TDP-43 Protein Fibril Formation Protocol

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TDP-43 PFFs were prepared as previously reported 11 (link), 12 (link). Monomeric TDP-43 (Proteintech, Wuhan, China) was resuspended in sterile water at concentration of 0.5 mg/ml. To obtain TDP-43 PFFs, the samples were incubated at 37 ºC for 2 hours with agitation at 600 rpm in an Eppendorf Thermomixer. For electron microscopy imaging, a 5 µl sample was absorbed onto carbon/formvar-coated 150 mesh copper grids (Yasheng Electronics Technology Co, Ltd., Zhengzhou, China) for 30 seconds and then stained with 5 µl of 2% uranyl acetate for 30 seconds. After staining, the grids were rinsed briefly in distilled water. The excess water was removed with filter paper, and the grids were air-dried prior to analysis by transmission electron microscopy (JEOL USA, Inc., Peabody, MA, USA).
Zhang R., Chen Y., Wang X., Tian H., Liu H., Xiang Z., Qi D., Huang J.H., Wu E., Ding X, & Wang X. (2021). Spreading of pathological TDP-43 along corticospinal tract axons induces ALS-like phenotypes in Atg5+/- mice. International Journal of Biological Sciences, 17(2), 390-401.
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2

Aerodynamic Particle Size Analysis

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Aerodynamic diameter size-distribution measurements were conducted within the VDT. These measurements were collected using an electrical low-pressure impactor (ELPI; Dekati, Tampere, Finland) and an aerodynamic particle sizer (APS, 3021 TSI Inc.). These measurements provided overlapping particle size distributions (by number) from 27 nm to 20 μm (ELPI, 27 nm to 10 μm and APS from 500 nm to 20 μm).
Three separate dispersions of 5 mg for each of the four materials were aerosolized within the VDT for the size-distribution measurements. These experiments used a sampler configuration described in Evans et al. (2010 (link), 2013 (link)) with all instruments sampling through a single sampling port connected to the VDT chamber through a 3-way valve that was also connected to a HEPA filter. Immediately following the material dispersion in the VDT, the 3-way valve was switched from sampling through the HEPA filter to sampling the aerosol generated in the VDT. All instruments were set to log data every second and data for the first 15 s of each dispersion were collected. During these size distribution experiments, samples were also collected using 25-mm styrene OFC samplers with mixed cellulose ester filters (0.8 μm pore size; SKC, Inc.) and analyzed by transmission electron microscopy (JEOL USA, Inc., Peabody, MA, USA) to examine particle morphology.
Dahm M.M., Evans D.E., Bertke S, & Grinshpun S.A. (2019). Evaluation of total and inhalable samplers for the collection of carbon nanotube and carbon nanofiber aerosols. Aerosol science and technology : the journal of the American Association for Aerosol Research, 53(8), 958-970.
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3

Ultrastructural Examination of NHBE Cells

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NHBE cells were digested with 0.25% trypsin to prepare single-cell suspension, centrifuged at 1000 rpm for 10 min, washed twice with prechilled PBS, and then fixed with prechilled 2.5% glutaric acid for 90 min at 4 °C. After washing in PBS, the cells were fixed with 1% osmium tetroxide at 4 °C for 30 min. After fixing, the cells were dehydrated with gradient ethanol and acetone, and then embedded in Epon812 resin (Sigma). The embedded blocks were cut into ultrathin sections using an ultramicrotome, and stained with uranyl acetate and lead citrate for ultrastructural examination under transmission electron microscopy (JEOL, Tokyo, Japan).
Yang B., Yang N., Chen Y., Zhu M., Lian Y., Xiong Z., Wang B., Feng L, & Jia X. (2021). An Integrated Strategy for Effective-Component Discovery of Astragali Radix in the Treatment of Lung Cancer. Frontiers in Pharmacology, 11, 580978.
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4

Nanoparticle Characterization by TEM

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After sonication, 20 μL of each test material suspension was placed on a carbon‐supported 200 mesh micro grid membrane (200‐Cu; Nisshin EM Co., Ltd, Tokyo, Japan); the shape of the nanoparticles was imaged by transmission electron microscopy (JEOL Co. Ltd, Tokyo, Japan), and the photos were analyzed by NIH image analyzer software (NIH, Bethesda, MD, USA). Over 1000 particles of each type of material were measured. Element analysis was done using a scanning electron microscope with an X‐ray microanalyzer (ADEX, Tokyo, Japan) after loading small aliquots of each test material directly after freeze‐drying on carbon conductive double‐faced adhesive tabs (Nisshin EM Co., Ltd).
Abdelgied M., El‐Gazzar A.M., Alexander D.B., Alexander W.T., Numano T., Iigou M., Naiki‐Ito A., Takase H., Abdou K.A., Hirose A., Taquahashi Y., Kanno J., Tsuda H, & Takahashi S. (2018). Potassium octatitanate fibers induce persistent lung and pleural injury and are possibly carcinogenic in male Fischer 344 rats. Cancer Science, 109(7), 2164-2177.
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5

Tight Junction Ultrastructure Analysis

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Cell monolayers were washed with PBS, gently scraped, and centrifuged at 1200 rpm. Cell pellets were fixed with the electron microscopy fixing buffer. Transmission electron microscopy (JEOL, Tokyo, JPN) was used to observe the tight junction ultrastructure of the ultrathin pellet sections.
Hou Z., Ding Q., Li Y., Zhao Z., Yan F., Li Y., Wang X., Xu J., Chen W., Wu G., Ruan X, & Zhao L. (2022). Intestinal epithelial β Klotho is a critical protective factor in alcohol-induced intestinal barrier dysfunction and liver injury. eBioMedicine, 82, 104181.
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6

Phage Morphology Characterization by TEM

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Phage aliquot with a titer of ~1011 pfu mL−1 was prepared for TEM observation and negatively stained with 4% w/v uranyl acetate (pH 7.2). The phage was observed using a JEOL transmission electron microscopy operated at 80 kV at the Electron Microscopy Laboratory in the University of Crete. From the obtained digital micrographs, capsid width and tail length of individual phages (n = 20) were measured with ImageJ software version 1.53p.
Tsertou M.I., Triga A., Droubogiannis S., Kokkari C., Anasi G, & Katharios P. (2023). Isolation and characterization of a novel Tenacibaculum species and a corresponding bacteriophage from a Mediterranean fish hatchery: Description of Tenacibaculum larymnensis sp. nov. and Tenacibaculum phage Larrie. Frontiers in Microbiology, 14, 1078669.
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7

Preparation and Characterization of G4Arg-DNA Complexes

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pOSKM was prepared by incorporating Oct4, Sox2, Klf4, and c-Myc genes as described previously.8 (link) To evaluate the transfection efficiency in vitro, the green fluorescent protein (GFP) gene was incorporated to produce pOSKMG.9 (link) The plasmids, in diethylpyrocarbonate (DEPC) water, were added to the G4Arg solution at various nanoparticle to DNA nitrogen-phosphorus (N/P) ratios (1:1, 5:1, 10:1, 15:1, 20:1, 35:1, and 50:1). After 10 minutes of mixing by vibration and then 20 minutes of sedimentation at room temperature, we obtained G4Arg-DNA complexes. The zeta potential and particle size of the G4Arg-DNA complexes in phosphate-buffered saline (PBS) were determined by a Zetasizer Nano (Malvern Instruments, Malvern, UK). Transmission electron microscopy (JEOL, Tokyo, Japan) was used for morphological characterization of the G4Arg-DNA complexes.
Zhu K., Li J., Lai H., Yang C., Guo C, & Wang C. (2014). Reprogramming fibroblasts to pluripotency using arginine-terminated polyamidoamine nanoparticles based non-viral gene delivery system. International Journal of Nanomedicine, 9, 5837-5847.
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8

Ultrastructural Analysis of Rat Brain Regions

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After the experiment was complete, rats were sacrificed by cervical dislocation, and prefrontal cortex and hippocampal tissue were cut according to the Rat Brain Stereotaxic Coordinates[69 ] on ice. The specimens were then cut into 1-mm3 blocks and fixed in 1% (v/v) osmium tetroxide fixative, then serially sliced into thin slices at 70 nm thickness for transmission electron microscopy (JEOL, Tokyo, Japan) observation.
Hou X., Zhang R., Lv H., Cai X., Xie G, & Song X. (2014). Acupuncture at Baihui and Dazhui reduces brain cell apoptosis in heroin readdicts. Neural Regeneration Research, 9(2), 164-170.
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9

Caveolae Structure Analysis in HUVECs

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Prior to electron microscopy assay, HUVECs were seeded at 1 × 106 cells/well in 6-well plates and grown overnight with adherent walls. Afterwards, the cell samples were treated with 0 and 250µM H2O2 for 30 min, respectively. Samples were collected and fixed with electron microscope stationary liquid, and stored at 4 °C for 3 days. Samples were washed three times with 0.1 M PBS, then soaked in a solution containing osmium tetroxide (1%) for 1 h, and then washed again with PBS. After a further staining in 1% aqueous uranyl acetate for 30 min, those samples were dehydrated and embedded in 50%, 70%, 90%, 100% ethanol and anhydrous acetone for 20 min, respectively. And then the cells were treated with a 1:1 volume mixture of anhydrous acetone and embedding agent and shaken for 2 h. Thereafter, the cells were treated with pure embedding agents and shaken for 2 h. The embedding agents were polymerized in an oven as the following conditions: 37 °C for 24 h, 45 °C for 24 h, and 60 °C for 48 h. A thin slice (120 nm) was made and the caveolae structure on the surface of HUVECs were observed by using transmission electron microscopy (80 kV, 15,000×; JEOL, Japan).
Zhao Y., Yu J., Huang A., Yang Q., Li G., Yang Y, & Chen Y. (2023). ROS impairs tumor vasculature normalization through an endocytosis effect of caveolae on extracellular SPARC. Cancer Cell International, 23, 152.
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10

Transmission Electron Microscopy Tissue Preparation

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Tissues were fixed with 2.5% glutaraldehyde for 30 min at room temperature, washed three times with 0.1 M phosphate buffer, fixed for 3 h in 1% osmium fixative, and then washed three more times. They were then dehydrated with graded concentrations of ethanol and different concentrations of acetone; after embedding, tissues were placed in an oven at 37°C to cure overnight, followed by 45°C for 12 h and 60°C for 48 h. Sections were double-stained with 3% uranyl acetate and lead citrate, and the slides were observed using transmission electron microscopy (JEOL, Japan).
Deng Y., Zhang Z., Yang H., Wang J., Feng L., Su Y, & Xu D. (2022). Mycophenolic Acid Induces the Intestinal Epithelial Barrier Damage through Mitochondrial ROS. Oxidative Medicine and Cellular Longevity, 2022, 4195699.
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