The electron microscopy was conducted at the Shanghai Jiao Tong University Instrumental Analysis Center. Briefly, the liver tissues from Cp-LKO and littermate controls were excised into pieces of 1 mm3 and fixed overnight in 2.5% glutaraldehyde at room temperature. The samples were post-fixed in potassium ferrocyanide-reduced osmium tetroxide (Electron Microscopy Sciences, USA) for 1.5 h and dehydrated in a bath with gradually increasing concentrations of ethanol (50%, 70%, 90%). The ethanol was then eliminated with gradually increasing acetone concentrations (90% ethanol: 90% acetone = 1:1, 90% acetone, 100% acetone). Further, the tissues were incubated with a mixture of acetone: EPON (liquid epoxy resin) (1:1,1:2, and 1:3) and ultimately embedded with pure EPON (Electron Microscopy Sciences, USA). The picture was captured using a biological transmission electron microscope (Tecnai G2 Spirit BioTWIN, Thermo Fisher Scientific, USA). The liver tissues from the BCS-treated Cp-LKO mice and the control group were analyzed following the same procedures.
Tecnai g2 spirit biotwin
Manufactured by Thermo Fisher Scientific
Sourced in United States, Germany, Netherlands, Czechia
The Tecnai G2 Spirit BioTWIN is a transmission electron microscope designed for biological samples. It provides high-resolution imaging capabilities for the analysis of cellular structures and molecular complexes.
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Lab products found in correlation
Em uc7, by Leica (8 mentions)
Veleta ccd camera, by Olympus (5 mentions)
Eagle 4k hs digital camera, by Thermo Fisher Scientific (5 mentions)
Uv 1601 pc, by Shimadzu (4 mentions)
Orius ccd camera, by Ametek (4 mentions)
Uct ultramicrotome, by Leica (4 mentions)
Em uc7 ultramicrotome, by Leica (4 mentions)
Zetaview s n 17 310, by Particle Metrix (3 mentions)
Ultracut uct ultramicrotome, by Leica (3 mentions)
Durcupan epoxy resin, by Merck Group (3 mentions)
Uc7 ultramicrotome, by Leica (3 mentions)
Nicolet 6700, by Thermo Fisher Scientific (3 mentions)
Potassium ferrocyanide reduced osmium tetroxide, by Electron Microscopy Sciences (2 mentions)
Pure epon, by Electron Microscopy Sciences (2 mentions)
Ultracut uct, by Leica (2 mentions)
Uranyl acetate, by Ladd Research Industries (2 mentions)
Formvar carbon coated copper grid, by Electron Microscopy Sciences (2 mentions)
Ftir 8201 pc infrared spectrophotometer, by Shimadzu (2 mentions)
Amt 2k ccd camera, by Advanced Microscopy Techniques (2 mentions)
Lambda 950, by PerkinElmer (2 mentions)
260 protocols using tecnai g2 spirit biotwin
1
Ultrastructural Analysis of Liver Tissues in Cp-LKO Mice
2
Transmission Electron Microscopy of sCABs
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The sCABs pellets were resuspended in paraformaldehyde (PFA, 2%). TEM (Tecnai G2 Spirit Bio TWIN, Thermo Fisher Scientific, USA) was applied to observe the morphology of sCABs stained with a uranyl‐oxalate solution (pH 7.0) and with methyl cellulose‐UA at 120 kV by EM.[45 ]
3
Characterization of Exosomes in Buruli Ulcer
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Exosomes isolated from the plasma samples of patients with active BU and healthy controls were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and WB.20 (link),21 (link) Exosomes were resuspended in PBS and then placed in an electron microscope grid stained with 2% uranyl acetate solution for 10 minutes at 25°C as described previously.22 (link) Exosomes were then identified using TEM (Tecnai G2 Spirit BioTWIN; Thermo Fisher Scientific, Waltham, MA, USA) (Fig. 1 A). The light-scattering and Brownian motion properties were utilized to calculate size distributions and exosome concentrations using NTA (Fig. 1 B). ZetaVIEW S/N 17-310 was used for the NTA (Particle Metrix, Meerbusch, Germany). Statistical analysis was carried out using the corresponding software (ZetaVIEW 8.04.02). Exosome markers including CD9, CD63, TSG101, and the calnexin protein were detected using WB analysis (described later) (Fig. 1 C).
4
Transmission Electron Microscopy of Plant Tissues
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Transmission electron microscopy (TEM) was performed as described by Zhao et al. with a minor modification [51 (link)]. The first true leaves were cut into small sections (1–2 mm in length) and then fixed in 2.5% glutaraldehyde under 4 °C overnight. The tissue samples were washed 3× in 0.1 M phosphate buffer and fixed in 2% osmic acid (OsO4) at 4 °C for 2 h. The tissues were washed three times with 0.1 M phosphate buffer and then dehydrated with an ethanol series of 50% v/v, 70% v/v, and 90% v/v for 15 min per ethanol concentration. The tissue samples were infiltrated with mixtures of 90% ethanol and 90% acetone (1:1), 90% acetone, and 100% acetone, acetone and resin (1:1, 1:2), and 100% resin for 12 h. The tissue samples were embedded and polymerized at 60 °C for 48 h. Thin sections (50–70 nm) were prepared with a cryo-ultramicrotome (UC6-FC6, Leica Microsystems, Wetzlar, Germany) and double-stained with uranyl acetate-lead citrate. The tissue samples were examined under a transmission electron microscope (Tecnai G2 Spirit BioTWIN, Thermo Fisher Scientific, Waltham, MA, USA).
5
Ultrastructural Analysis of Mouse Ileal Tissue
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Transmission electron microscopy was used to visualize ultrastructural changes in mouse ileal tissue. Therefore, ileal tissues were initially fixed overnight using 2.5% glutaraldehyde buffered in cacodylate. The embedding procedure comprised fixation in 1% osmium tetroxide, dehydration in a graded ethanol series with an incubation step with uranyl acetate (between the 50 and 90% ethanol step), and finally rinsing in propylene oxide. The specimens were then embedded in epoxy resins that polymerized for 16 h at 60°C. After embedding, semithin sections (0.5 µm) were cut using an ultra-microtome (Ultracut UCT; Leica Biosystems) with a diamond knife. Sections were stained with Toluidine blue, placed on glass slides, and examined by light microscopy to select appropriate areas for ultrathin preparation. Ultrathin sections (50–70 nm) were cut again using an ultra-microtome. Sections were mounted on copper grids and contrasted with uranyl acetate for 2–3 h at 42°C followed by lead citrate for 20 min at room temperature. Imaging was performed using a transmission electron microscope (Tecnai G2 Spirit Biotwin; Thermo Fisher Scientific) at an operating voltage of 120 kV.
6
Negative Staining of S-CREM1 Nanoparticles
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The carbon-coated copper grids (200-mesh) were subjected to glow discharge for 20 s. Ten microliters of purified S-CREM1 suspensions were adsorbed to carbon-coated copper grids for 5 min and negatively stained twice with 2% (w/v) uranyl acetate for 10 s and 30 s, respectively. The stained sample was dried for 10 min and observed using a Tecnai G2 Spirit BioTwin transmission electron microscope (FEI Tecnai G2 F20, Thermo Fisher Scientific, Waltham, MA, USA).
7
Characterization of GPEI Polyplexes
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The particle size and zeta potential of GPEI polyplexes at various w/w ratios were measured by Particle Size Analyzer (Brookhaven Particle Size Analyzer 90 Plus, Brookhaven Instruments Corporation, Holtsville, NY, USA), and PEI 25 kDa polyplexes (w/w ratio = 2) were used as the positive control. The morphology of GPEI polyplexes at w/w ratio of 5 was examined by transmission electron microscopic (TEM) (Tecnai G2 spirit Biotwin, Thermo Fisher Scientific, Waltham, MA, USA).
8
Autophagosome Formation in HLECs via TEM
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The formation of autophagosomes in HLECs was observed by transmission electron microscopy (TEM) as previously described (23 (link)). Briefly, HLECs grown on slides were fixed for 1 h with 2.5% glutaraldehyde (cat. no. G7776; Sigma-Aldrich; Merck KGaA) dissolved in 0.1 mol/l PBS at room temperature, incubated with 1% osmium tetroxide (cat. no. O5500; Sigma-Aldrich; Merck KGaA) for 30 min at room temperature and observed using a TecnaiG2 Spirit Bio TWIN transmission electron microscope (120 kV; FEI; Thermo Fisher Scientific, Inc.). The TEM experiments were performed at least three times.
9
Characterizing Influenza M2 Protein VLPs
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3FLAG-M2-WT or 3FLAG-M2-E79K were transfected on 293T cells to generate VLPs. Twenty-four hours post transfection, supernatants were collected and centrifuged at 2000× g for 20 min to remove the cell debris. MDCK cells were infected with CA09-M2-WT or CA09-M2-E79K viruses; 48 h post inoculation the supernatant was collected and centrifuged at 2000× g for 20 min to remove the cell debris. The clarified supernatants were ultra-centrifuged at 20,000× g and cushioned with 20% (w/v) sucrose. After centrifugation, the pellets were collected on ice; the virus particles were fixed in 4% paraformaldehyde at 37 °C overnight and then sent for electric microscopy. The service was provided by the Instrumental Analysis Center at Shanghai Jiao Tong University with Tecnai™ G2 spirit Biotwin (Thermo Fisher, Grand Island, NE, USA).
10
Ultrastructural Analysis of Eye Tissues
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