A small portion (∼1 mm3) of the hypothalamus from one mouse in each group was sectioned and incubated for 2 h at 4°C in 2.5% glutaraldehyde (Solarbio Life Science, P1126, Beijing, China). The specimens were rinsed with 0.1 M phosphoric acid, postfixed in 1% osmium tetroxide for 2–3 h, and then rinsed with 0.1 M phosphoric acid again for 15 min × 3 times. The specimens were dehydrated with different concentrations of ethanol at 4°C, and put into acetone three times at room temperature, then embedded in epoxy resin. After curing in the oven, the specimen is cut into ultrathin sections of 50–60 nm which were stained with lead citrate and examined by transmission electron microscopy (JEM-1010, Japan). Then we randomly selected any cell in the hypothalamus tissue for observation, with 10 visual fields for each cell and counted the number of autophagosomes in the 10 visual fields of each group by observing the electron microscope pictures.
P1126
Manufactured by Solarbio
Sourced in China
The P1126 is a laboratory equipment designed for the purpose of pH measurement. It features a digital display and provides accurate pH readings.
Automatically generated - may contain errors
Lab products found in correlation
Tecnai 20, by Philips (2 mentions)
Ht7800, by Hitachi (2 mentions)
Ethylene diamine tetraacetic acid (edta), by Solarbio (1 mentions)
B53 microscope, by Olympus (1 mentions)
Ab pas staining kit, by Solarbio (1 mentions)
Jem 2100 plus, by JEOL (1 mentions)
Formvar coated grids, by Ted Pella (1 mentions)
Jem 2100, by JEOL (1 mentions)
Fei tecnai g2 f30, by Thermo Fisher Scientific (1 mentions)
Nova nanosem 450, by Thermo Fisher Scientific (1 mentions)
Leica uc7, by Leica camera (1 mentions)
Hy 100579, by MedChemExpress (1 mentions)
Osmium tetroxide, by Merck Group (1 mentions)
Em uc7, by Leica camera (1 mentions)
Jem 1400, by JEOL (1 mentions)
Epon 812 resin, by TAAB (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Alkaline lead citrate, by Merck Group (1 mentions)
O5500, by Sinopharm (1 mentions)
Jem 1230 tem, by JEOL (1 mentions)
22 protocols using p1126
1
Quantification of Autophagosomes in Mouse Hypothalamus
2
Ultrastructural Analysis of Corpus Callosum
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After the mice were deeply anesthetized, the brain tissue was quickly removed, and the corpus callosum was separated from the ice. The tissue was immediately fixed in 2.5% glutaraldehyde (P1126; Beijing Solarbio Science & Technology Co., Ltd.), and a 1‐mm cube tissue block was obtained from the corpus callosum of the mice. Then, the tissue was quickly placed in 2.5% glutaraldehyde for incubation at 4°C overnight, and then fixed with 2% osmium tetroxide in PBS. After dehydration through an alcohol gradient, the samples were embedded in epoxy resin. Semi‐thin sections were then examined by transmission electron microscopy. The G‐ratio, which refers to the diameter of the axon/the diameter of the entire myelinated fiber, is used as an indicator.
34 (link)
34 (link)
3
Ultrastructural Analysis of Extracellular Vesicles
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To detect the morphological structure of MSCs-EVs and MRC5-EVs, a drop of EVs (1 μg/μL) was transferred onto a carbon film (Zhongjingkeji Technology, Beijing, China) with negative staining by 2% phosphotungstic acid. Next, transmission electron microscopy (TEM) images of MSCs-EVs and MRC5-EVs were captured from air-dried samples by the TEM (Talos F200C, MA, USA).
Preparation of mouse heart samples for TEM has been reported as previously described [31 (link)]. Briefly, excised hearts were cut into small tissue blocks (1 mm3) and fixed with 2.5% glutaraldehyde (Solarbio, P1126, Beijing, China) at 4 °C for 12–24 h and rinsed with PBS 3 times. Then, samples were postfixed in 2% osmium tetroxide (Sigma-Aldrich, 208,868, St Louis, MO, USA) for around 2 h, dehydrated in alcohol and infiltrated overnight with epoxy resin. Finally, after air dry for 48 h at 60 °C, samples were cut into an ultrathin section at 60 nm by an ultramicrotome (Reichert Ultracut, Vienna, Austria) followed by staining with uranyl acetate and lead citrate. The microstructure of ER in ventricular myocytes was captured by TEM.
Preparation of mouse heart samples for TEM has been reported as previously described [31 (link)]. Briefly, excised hearts were cut into small tissue blocks (1 mm3) and fixed with 2.5% glutaraldehyde (Solarbio, P1126, Beijing, China) at 4 °C for 12–24 h and rinsed with PBS 3 times. Then, samples were postfixed in 2% osmium tetroxide (Sigma-Aldrich, 208,868, St Louis, MO, USA) for around 2 h, dehydrated in alcohol and infiltrated overnight with epoxy resin. Finally, after air dry for 48 h at 60 °C, samples were cut into an ultrathin section at 60 nm by an ultramicrotome (Reichert Ultracut, Vienna, Austria) followed by staining with uranyl acetate and lead citrate. The microstructure of ER in ventricular myocytes was captured by TEM.
4
Ultrastructural Analysis of Bone Tissue
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The traction tissue from the experimental side and normal bone tissue from the CS were trimmed into tissue blocks of 1 mm3 for transmission electron microscopy (TEM) (JEM1200EX, Japan). The sections were fixed in 2.5% glutaraldehyde at 4°C for 24 to 48 hours (P1126; Solarbio, China). EDTA (E1171; Solarbio) was used at 4°C for 2 weeks (with weekly changes of EDTA) to decalcify bone. Acetone was used to dehydrate tissue blocks for 30 minutes. Ultrathin sections (700–900 nm) were stained with uranium hydrogen acetate and lead citrate and observed under TEM.
5
Histological and Ultrastructural Analysis of Biological Samples
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Briefly, HE and PAS samples were fixed using 4% paraformaldehyde, and 8 µm thick sections were prepared. Sections were dewaxed with xylene and ethanol and stained with HE and PAS with using a hematoxylin and eosin staining kit (G1120; Solarbio, China) and AB-PAS staining kit (G1285; Solarbio). For transmission electron microscopy (TEM) samples, fixation was performed using 2.5% glutaraldehyde (P1126; Solarbio) and 1% osmic acid. After dehydration using a gradient of ethanol and acetone, the samples were embedded in epoxy resin. Subsequently, 70 nm-thick sections were prepared using an EM UC7 ultrathin sectioning machine (Leica, Germany). Finally, sections were stained with 3% uranyl acetate-lead citrate. HE and PAS-stained sections were imaged using a B×53microscope (Olympus, Japan), and TEM-stained sections were imaged using a JEM-2100 Plus (JEOL, Japan).
6
Ultrastructural Analysis of CL in Pregnant Rats
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CL tissues were collected from ovaries of pregnant rats with or without 3-MA treatment. Specimens for TEM were prepared and fixed with 2.5% glutaraldehyde (P1126, Solarbio) in PBS (4 °C, pH 7.4, 0.1 M) for 24 h. The samples were then postfixed with 1% OsO4 (Ted Pella) for 1.5 h and stained with 3% aqueous uranyl acetate for 1 h after washing. After that, the samples were dehydrated by graded alcohol series, embedded in Araldite (90529-77-4, SPI-CHEM), sectioned with a thickness of approximately 60 nm and mounted on Formvar-coated grids (01700-F, Ted Pella). The ultrathin sections were contrasted with 0.3% lead citrate stainning, examined and photographed under a transmission electron microscope (JEM-2100, JEOL, Tokyo, Japan).
7
Transmission Electron Microscopy Sample Preparation
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A 6-well plate of cell cultures was fixed for 24 h with a solution (P1126, Solarbio, Beijing) containing 2.5% glutaraldehyde in 0.1 mM phosphate buffer saline (PBS). The cells were washed in 0.1 M PBS, fixed with 1% buffered osmium, and stained with 1% Millipore-filtered uranyl acetate. The samples were incubated at 60 °C for 48 h following dehydration and embedding. Transmission electron microscope images were taken finally.
8
Transmission Electron Microscopy of Cells
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U251 cells were seeded in the 10 cm dish at a density of 1 × 106, and then exposed to nanomaterials for 24 h. After being digested by trypsin, cells were centrifuged at 500 × g for 5 min into clumps. Subsequently, cells were fixed in 2.5% glutaraldehyde (P1126, Solarbio, Beijing, China) (diluted in 0.1 µM PBS; pH 7.4) at 4 °C for 24 h and then post-fixed in 1% osmium tetroxide (201030, Merck, New Jersey, USA) (dissolved in PBS; pH 7.4) at 25 °C for 60 min. After dehydration using different concentrations of Ethanol (30%, 50%, 80%, 90%, 100%), samples were embedded by resin (45347, Merck, New Jersey, USA) with different conditions (37 °C for 12 h; 45 °C for 12 h; 60 °C for 12 h) and ultrathin sectioning (the thickness is 50 nm), the samples were stained with uranyl acetate at RT for 60 min and stained with lead citrate (15326, Merck, New Jersey, USA) at RT for 8 min. Digital images were captured using TEM (FEI Tecnai G2 F30; Thermo Fisher Scientific, Inc.).
9
Ultrastructural Characterization of AMEVs
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AMEVs (1 uL) were resuspended in 100uL PBS, and then resuspended in 900uL glutaraldehyde solution (2.5%) (P1126, BeiJing Solarbio Science & Technology Co. Ltd, China) at 4 °C overnight. Micrographs of AMEVs were obtained using a low vacuum scanning electron microscopy (Nova Nano SEM 450, FEI, USA) operated at 5 KV.
10
Ultrastructural Analysis of SSC Mitochondria and Autophagosomes
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TEM was used to characterize the morphology and structure of SSC mitochondria and autophagosomes. Briefly, the frozen-thawed SSCs were washed with PBS and centrifuged to remove the supernatant. The cells were fixed with 2.5% glutaraldehyde (P1126; Solarbio) for 4–6 h and then fixed with 1% osmium tetroxide for 2–4 h. Next, the samples were dehydrated in graded alcohol and embedded in Epon. The prepared specimens were ultrathin sectioned and added to the copper grid. TEM (Hitachi, Japan) was used to visualize changes in the ultrastructure of SSCs, with 15 or 12 cells per group.
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