MSCs were seeded in 24-well plates at a density of 7 × 104 cells/well. At 24 h after plating, MSCs were exposed to Printex 90 at concentrations of 0, 10 μg/mL for 24 h. After trypsinization, a cell pellet was fixed with 2% glutaraldehyde. Cells were imbedded, cut into ultrathin slices, and viewed using a TEM (FEI Tecnai G2 Spirit Bio TWIN, USA).
Tecnai g2 spirit bio
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Tecnai G2 Spirit Bio is a high-performance transmission electron microscope (TEM) designed for biological applications. It provides high-resolution imaging and advanced analytical capabilities for the study of biological samples.
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5 protocols using tecnai g2 spirit bio
1
Transmission Electron Microscopy of MSCs
2
Transmission Electron Microscopy of Leaf Samples
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The Gy14 and yf leaf samples were collected from the 14th true leaves for TEM. Leaf samples of the 9930 WT and CsSRP43 CRISPR transgenic lines were collected from the second expanded leaf. These leaves were cut into small pieces of ~0.2 mm × 0.2 mm size and fixed in a 4.0% glutaraldehyde solution. Leaf samples were prepared for TEM according to Xiong et al. [67 (link)]. Finally, the samples were observed using a Tecnai G2 Spirit Bio (FEI, USA) transmission electron microscope.
3
Spinal Cord Injury Histological Analysis
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Areas 1 mm above and below the lesion area of the spinal cord were isolated and embedded in paraffin dividing into 3 μm thick sections. We isolated areas 1 mm above and below the lesion area of the spinal cord and embedded them in paraffin dividing them into 3 μm thick sections. The sections were stained with luxol fast blue (LFB) and sealed with neutral resin. The images were observed under microscopy (Olympus, Tokyo, Japan). Six of 30 serial paraffin sections were analyzed in each analysis. Electron microscopy was carried out on spinal cord sections of SCI mice at 28 dpi. Briefly, spinal cords were perfused with 3% paraformaldehyde and 1% glutaraldehyde and samples were observed at 70 nm by transmission electron microscope (FEI Tecnai G2 Spirit Bio TWIN, NY, USA). For G-ratio analysis, at least 100 fibers of each mouse were measured [30 (link)].
4
Mitochondrial Ultrastructure Analysis
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Mouse hearts were fixed in 4% glutaraldehyde, postfixed in 1% osmium tetroxide, dehydrated in gradient acetone, and embedded in Epon resin. The resin was then polymerized at 30 °C, 45 °C, and 60 °C for 1 day per temperature. The embedded tissues were cut at 70 nm, and the sections were stained with uranyl acetate and lead citrate. Transmission electron microscopy (FEI Tecnai G2 Spirit Bio TWIN, Japan) was used to observe the mitochondrial ultrastructure.
5
Transmission Electron Microscopy of Exosomes
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The identified HFSCs were fixed with 2.5% glutaric PBS for 4 h or overnight. Thereafter, the cells were rinsed with PBS, fixed with 1% osmium tetroxide, rinsed with double-distilled water (ddH2O), fixed/stained with 2% uranyl acetate, and gradient dehydrated in 50%, 70%, 90%, 100% ethanol, and acetone, prior to infiltration, embedding, and polymerization. Sections of the embedded cells were cut using an ultramicrotome, followed by staining with uranyl acetate and lead citrate.
Isolated exosomes derived from DPCs were diluted 10 times, fixed in 4% paraformaldehyde, and then layered on a carbon/Formvar film-coated TEM grid (EMCN®, Beijing, China) for 20 min. The grids were treated with 1% glutaraldehyde and then washed eight times with ddH2O (2 min each time), and stained with 1% uranyl acetate for 10 min. The preparations were dried naturally and observed using a transmission electron microscope (Tecnai G2 Spirit Bio, FEI Company, Hillsboro, OR, USA). The Feret's diameter of the particles was determined using ImageJ software (Rasband, W.S., U.S. National Institutes of Health, Bethesda, Maryland, USA,http://imagej.nih.gov/ij/ , 1997-2014).
Isolated exosomes derived from DPCs were diluted 10 times, fixed in 4% paraformaldehyde, and then layered on a carbon/Formvar film-coated TEM grid (EMCN®, Beijing, China) for 20 min. The grids were treated with 1% glutaraldehyde and then washed eight times with ddH2O (2 min each time), and stained with 1% uranyl acetate for 10 min. The preparations were dried naturally and observed using a transmission electron microscope (Tecnai G2 Spirit Bio, FEI Company, Hillsboro, OR, USA). The Feret's diameter of the particles was determined using ImageJ software (Rasband, W.S., U.S. National Institutes of Health, Bethesda, Maryland, USA,
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