BG11 medium was purchased from Freshwater Algae Culture Collection at the Institute of Hydrobiology (Wuhan, Hubei, China). Bismuth (powder, 325 mesh, 99.5%) was purchased from Alfa-Aesar Co., Ltd (Shanghai, China). Roswell Park Memorial Institute (RPMI) 1640 medium, High glucose DMEM medium, fetal bovine serum (FBS), phosphate buffer saline (PBS), penicillin-streptomycin and trypsin enzyme were purchased from Thermo Fisher Scientific (Waltham, MA, USA). [Ru(dpp)3]Cl2 was obtained from Alfa-Aesar Co. Ltd (Shanghai, China). Cell Counting Kit-8 (CCK-8) was bought from Dojindo Laboratories (Tokyo, Japan). Crystal Violet Staining Solution and 2′,7′-dichlorofluorescin diacetate (DCFH-DA) was purchased from Beyotime (Shanghai, China). 4% Paraformaldehyde, DAPI stain solution and bovine serum albumin (BSA) were purchased from Sangon Biotech (Shanghai, China). Triton X-100 was obtained from Diamond (Shanghai, China). γ-H2AX antibody were purchased from Cell Signaling technology (Danvers, MA, USA). 2.5% glutaraldehyde fixative was obtained from Servicebio (Wuhan, Hubei, China). Anti-hypoxia-inducible factor 1alpha (HIF-1α) antibody was purchased from Santa Cruz Biotechnology (Dallas, Texas, USA). Annexin V-FITC/PI apoptosis kit was obtained from Multi Sciences (Hangzhou, Zhejiang, China). All chemical reagents were used directly without further depuration.
Glutaraldehyde fixative
Manufactured by Wuhan Servicebio Technology
Sourced in China
2.5% glutaraldehyde fixative is a laboratory reagent used for the preservation and fixation of biological samples. It functions as a cross-linking agent, helping to maintain the structural integrity of cells and tissues during preparation for analysis or storage.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Ht7800 ht7700, by Hitachi (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
1640 medium, by Thermo Fisher Scientific (1 mentions)
Phosphate buffer saline (pbs), by Thermo Fisher Scientific (1 mentions)
Trypsin enzyme, by Thermo Fisher Scientific (1 mentions)
Annexin 5 fitc pi apoptosis kit, by MultiSciences Biotech (1 mentions)
Paraformaldehyde (pfa), by Sangon (1 mentions)
γh2ax antibody, by Cell Signaling Technology (1 mentions)
Dmem high glucose medium, by Thermo Fisher Scientific (1 mentions)
Cell counting kit 8 cck 8, by Dojindo Laboratories (1 mentions)
Crystal violet staining solution, by Beyotime (1 mentions)
2 7 dichlorofluorescin diacetate dcfh da, by Beyotime (1 mentions)
Bovine serum albumin (bsa), by Sangon (1 mentions)
Dapi stain solution, by Sangon (1 mentions)
Beem capsules, by Electron Microscopy Sciences (1 mentions)
Leica uc7, by Leica camera (1 mentions)
Transmission electron microscope, by Hitachi (1 mentions)
4 protocols using glutaraldehyde fixative
1
Evaluating Bismuth Nanoparticle Cytotoxicity
2
Transmission Electron Microscopy of Cells
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HeLa cells were inoculated in 6-well plates transfected with the target plasmids for 36 h, or MARC-145 cells were infected with PRRSV strain HN07-1 for 12 h. The cells were collected and fixed using 2.5% glutaraldehyde fixative (Servicebio, Cat. No. G1102) at RT for 30 min. The fixed cells were dehydrated with a graded series of ethanol and finally embedded in pure LR White resin at 4°C (HaideBio, Cat. No. 14381-UC) using BEEM capsules (Electron Microscopy Sciences, Cat. No. 69920–00). After polymerization with a low-temperature UV polymerizer (UVCC2515; Electron Microscopy, Beijing, China), the resin blocks were sliced into 70- to 80-nm thin slices on an ultramicrotome (UC7; Leica, Wetzlar, Germany), and the ultrathin sections were then fished out onto the 150-mesh nickel grids with a Formvar film (Head Biotechnology Co., Cat. No. FCF200-Cu-50). Finally, the samples were observed at 80 kV using TEM (Ht7800/Ht7700; Hitachi, Tokyo, Japan). Representative images were shown.
3
Transmission Electron Microscopy Sample Preparation
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The collected cells were fixed in 2.5% glutaraldehyde fixative (Servicebio, China) for 30 minutes, followed by PBS cleaning and wrapping in 1% agarose. Then, fix at room temperature with 1% osmium acid in the dark for 2 hours, and sequentially add 30%, 50%, 70%, 80%, 95%, 100%, and 100% alcohol to dehydrate for 20 minutes each time, and 100% acetone twice, each time for 15 minutes. Then, after penetration embedding, remove the resin block and slice it into ultra-thin sections using an ultra-thin slicer (Leica UC7, Leica). The slices are then placed in a copper mesh box and dried overnight at room temperature. Finally, we observed and photographed under a transmission electron microscope (HT7800/HT7700, HITACHI).
4
Ultrastructural Analysis of Cultured Cells
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Cells (2 × 105 cells/well) seeded in a 6-well plate were cultured in DMEM and 10% FBS containing 10 mM glutamate and 25 µM SB202190 for 24 hours in a 37°C, 5% CO2 incubator. The cells were fixed with 2.5% glutaraldehyde fixative (Servicebio, Wuhan, China) at room temperature in the dark for 5 minutes. The cells were scraped off with a cell scraper and centrifuged (100 × g, 5 minutes). The fixative was discarded and fresh electron microscope fixative was added (Servicebio, Wuhan, China). The cells were fixed at room temperature for 30 minutes in the dark. Next, 1% osmium tetroxide (Servicebio) was used for post-fixing at room temperature (20°C) for 2 hours. After dehydration with gradient concentrations of alcohol (50%, 70%, 80%, 90%, and 100%) and acetone, cells were embedded in epoxy resin (Servicebio). The samples were cut into 60–80 nm ultra-thin sections for uranium-lead (Servicebio) double staining and then examined under a transmission electron microscope (Hitachi, Tokyo, Japan).
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