To perform transmission electron microscopy, 1 mm3 hippocampus DG region tissues were fixed overnight in electron microscope fixation solution (Servicebio, China) and dehydrated with 1% osmium tetroxide (v/v) for 2 h. After acetone penetration, the tissues were stained with 4.8% uranyl acetate, rinsed, and sliced with a lead-uranium double dyed copper mesh. The microphotographs were acquired using a high-resolution transmission electron microscope (JEOL, Japan).
Electron microscope fixation solution
Manufactured by Wuhan Servicebio Technology
Sourced in China
The Electron Microscope Fixation Solution is a chemical solution used to prepare biological samples for imaging under an electron microscope. It helps preserve the structural integrity of the sample by chemically fixing and stabilizing the cellular components. This solution is an essential tool for researchers and scientists conducting electron microscopy studies.
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5 protocols using electron microscope fixation solution
1
Transmission Electron Microscopy of Hippocampus DG
2
Liver Tissue Preparation for SEM
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The liver tissues were put into the electron microscope fixation solution (Servicebio) immediately after isolation and cut into small blocks about 2 mm × 2 mm × 2 mm. The tissues were stored at room temperature for 4 h away from light and transferred to 4°C for storage. After dehydration, drying, and gold-coated treatment, the prepared specimens were detected by an SEM (SU8010, Hitachi, Japan).
3
Ultrastructural Analysis of Cells
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Cells were harvested and fixed at 4 °C for 2–4 h with electron microscope fixation solution (Servicebio, China) and further fixed with 1% osmium in·0.1 M phosphate buffer for 2 h. Cells were dehydrated in graded ethanol (50%, 70%, 80%, 90%, 95% and 100%) and 100% acetone twice. Cells were embedded and cut into slices of 60–80 nm thickness with an ultra-thin slicing machine. Then, the sections were double-stained with 2% uranium acetate-lead citrate. Finally, photographs of eight random fields were captured for each sample under a transmission electron microscope (HITACHI, Tokyo, Japan).
4
SEM Analysis of Hepatic Sinusoid
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Freshly collected specimens were immediately immobilized in electron microscope fixation solution (Wuhan Servicebio Technology Ltd., Wuhan, China) for 2 hours and then transferred to 4 °C. The tissues were immobilized in 1% osmic acid buffer for 2 hours, serially dehydrated in ethanol and dried in a critical point drier (K850; Quorum Technologies Ltd, Lewes, United Kingdom). Then images were obtained under a scanning electron microscope (SU8010; Hitachi, Tokyo, Japan) after conductive treatment using an ion sputtering analyzer (MSP-2S; IXRF Sytems, Inc., Austin, TX, USA). The sections were observed by SEM to analyze the fenestrated structure of the hepatic sinus endothelium.
5
Isolation and Characterization of EPC-Derived Microvesicles
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EPCs were cultured for 7 days as previously described (30 (link)), washed twice with PBS and serum-starved for 12 h. Subsequently, MEM/F12 medium containing cultured EPCs was collected and centrifuged at 4°C (1,000 × g; 15 min), and the supernatant was extracted at 4°C (100,000 × g; 60 min) for the collection of secreted EPC-MVs. After embedding in Pon 812 epoxy resin (Structure Probe, Inc.), it was placed at 37°C overnight. Then, electron microscope fixation solution (Wuhan Servicebio Technology Co., Ltd.) was added for 4 h, and uranium acetate (2%; Wuhan Servicebio Technology Co., Ltd.) and lead citrate (Wuhan Servicebio Technology Co., Ltd.) were stained at 25°C for 15 min. MVs were confirmed via transmission electron microscopy. In the co-culture system, 50 µg/ml EPC-MVs (31 (link)) was added to the top chamber of a Transwell assay plate, and PRKs were added to the bottom chamber as previously described, and incubated for 24 h.
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