Cells were cultured in the osteogenic-/adipogenic-inducing medium to confirm the capability of multidirectional differentiation. The components of the inducing medium were described in our previous research [21 (link)]. After induction on day 21, 4% paraformaldehyde was used to fix cells. Then, Alizarin Red Staining kit (Sigma-Aldrich, St. Louis, USA) and Oil Red O kit (Solarbio, Beijing, China) were implemented to stain cells following the instructions of the manufacturers.
Oil red o kit
Manufactured by Solarbio
Sourced in China, United States
The Oil Red O kit is a laboratory reagent used for the staining and visualization of lipids in biological samples. The kit contains the necessary components, including the Oil Red O dye solution, to perform this specific staining procedure. The core function of the kit is to facilitate the detection and analysis of lipid content in cells or tissues.
Automatically generated - may contain errors
Lab products found in correlation
Inverted microscope, by Zeiss (2 mentions)
Alizarin red staining kit, by Merck Group (1 mentions)
Eclipse upright microscope, by Nikon (1 mentions)
Hematoxylin, by Solarbio (1 mentions)
Cm1950, by Leica (1 mentions)
Ix71 microscope, by Olympus (1 mentions)
Propidium iodide solution, by Solarbio (1 mentions)
Taqman microrna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
Annexin 5 fitc solution, by Solarbio (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Cell counting kit 8 cck 8, by Keygen Biotech (1 mentions)
11 protocols using oil red o kit
1
Multilineage Differentiation Evaluation
2
Lipid Droplet Staining in Stem Cells
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As per the instructions of Oil Red O kit (G1262, Solarbio, Beijing, China). The SCs were fixed with fixative for 20–30 min and washed. Then, the SCs were dyed with ready-to-use staining for 10–20 min, followed by cleaning with 60% isopropanol to clear the background. The nuclei were stained with Hematoxylin (G1140, Solarbio, Beijing, China). After washing with distilled water, cells were observed using an Eclipse upright microscope (Nikon, Tokyo, Japan).
3
Frozen Liver Tissue Staining
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The liver was placed in an embedding mold, covered with SAKURA Tissue-Tek® O.C.T. Compound (4583, SAKURA, Torrance CA, USA), wrapped in tin paper, and placed on the surface of liquid nitrogen for cooling and coagulation. After complete coagulation, slices that were 8 μm each were made using a frozen section machine (CM1950, Leica, Nussloch, Germany). According to the instructions of the modified Oil Red O kit (G1262, Solarbio, Beijing, China), the slides with adsorbed liver tissue sections were placed in 60% isopropanol for 2 min, taken to ice water (ice water mixture) for slight cleaning, put into the Oil Red O staining solution for 10 min (away from light), put into 60% isopropanol for 5 s, and washed with ice water; finally, hematoxylin staining was used to stain the nucleus for 5 min, and the edge of the tissue was dried after ice water washing. Glycerin gelatin was used to seal the film, and photographs were obtained under an inverted microscope.
4
Oil Red O Staining of Lipid Droplets
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Oil-Red-O staining was performed with Oil Red O Kit (G1262, Solarbio). Cells were washed with PBS for twice, and fixed with the fixative buffer for 30 min. Wash the cells with distilled water twice and then incubate in 60% isopropanol for 5 min. The newly prepared oil red O staining solution was added and soaked for 20 min. Mayer hematoxylin staining solution was added for 2 min. Discard the dye and wash it for 3 times. Oil-Red-O staining pictures were taken using an Olympus IX71 microscope.
5
Oil Red O Staining for Lipid Analysis
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Oil Red O staining was performed using Oil Red O kit (Solarbio, Beijing, China). Frozen sections were prepared, dried at room temperature for 15–20 min, incubated with 100% isopropanol for 5 min, and then incubated with 0.5% oil red O solution for 7–8 min; they were then washed with 85% isopropanol solution for 3 min. We dehydrated and washed; finally, we used mounting tablets to mount the slides and perform a microscopic examination.
6
Investigating the Effects of BMP7-Enriched sEVs on aHSCs
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The aHSCs (HSC-T6 and LX-2 cells) were divided into a control group (0 µg/mL sEVs), blank-sEVs group (100 µg/mL blank-sEVs), negative-sEVs group (100 µg/mL negative-sEVs), and BMP7+sEVs group (100 µg/mL BMP7+sEVs). After 48 h of the corresponding treatments, the cells in each group were scraped with a cell scraper before the samples were collected by centrifugation at 2000 × g for 3 min at 25 °C. After discarding the fixative, a new electron microscopy fixative was added. After cleaning, dehydration, resin infiltration, and embedding the sections in pure resin, the cells in each group were imaged using TEM after immunolabeling with colloidal gold.
Similarly, the aHSCs (HSC-T6 and LX-2 cells) were treated with 0 µg/mL sEVs, 100 µg/mL blank-sEVs, 100 µg/mL negative-sEVs, or 100 µg/mL BMP7+sEVs for 48 h before being stained with an Oil Red O kit (Solarbio). Specimens were observed and photographed under a microscope (Leica, Wetzlar, Germany) to detect red staining indicating the formation of lipid droplets.
Similarly, the aHSCs (HSC-T6 and LX-2 cells) were treated with 0 µg/mL sEVs, 100 µg/mL blank-sEVs, 100 µg/mL negative-sEVs, or 100 µg/mL BMP7+sEVs for 48 h before being stained with an Oil Red O kit (Solarbio). Specimens were observed and photographed under a microscope (Leica, Wetzlar, Germany) to detect red staining indicating the formation of lipid droplets.
7
Hepatic Steatosis Evaluation via Oil Red O
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To evaluate hepatic steatosis, 10‐μm frozen sections of the liver were stained with Oil Red O kit (Solarbio Science & Technology Ltd., Beijing, China).
8
Evaluating Lipid Accumulation in HCC Cells
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We treated HCC cells with verteporfin of different concentration gradient and cells were fixed under conventional six-well plate and subjected to oil red O staining after 48 hours of treatment. The non-esterified Free Fatty Acids (NEFA) was estimated in plasma by NEFA Colorimetric Assay Kit (E-BC-K013-M). NEFA ELISA kit (E-BC-K013-M) was purchased from Elabscience Biotechnology. Oil-Red-O staining was performed with Oil Red O Kit (G1262, Solarbio). Cells were washed with PBS for twice, and fixed with the fixative buffer for 30 min. Wash the cells with distilled water twice and then incubate in 60% isopropanol for 5 min. The newly prepared oil red O staining solution was added and soaked for 20 min. Mayer hematoxylin staining solution was added for 2 min. Discard the dye and wash it for 3 times. Oil-Red-O staining pictures were taken using a Zeiss inverted microscope. The Oil Red O Staining experiment was performed at least three independent repeat experiments. Quantifications were performed using Image J software.
9
Preadipocyte Lipidosis Quantification
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An Oil Red O Kit (Solarbio, USA) was used to stain porcine preadipocytes according to the manufacturer’s instructions. The morphology of lipid droplets after oil red O staining was observed under a microscope. A triglyceride kit (Applygen, China) was used to measure preadipocyte lipidosis following the manufacturer’s recommended protocol.
10
Oil Red O Staining of 3T3-L1 Adipocytes
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The Oil Red O Kit (Solarbio, Beijing, China) was used to stain the 3T3-L1 adipocytes according to the manufacturer’s instructions. Differentiated and mature 3T3-L1 adipocytes were taken, fixed with 4% paraformaldehyde for 20 min after being washed, immersed in and washed with 60% isopropanol for 5 min, immersed in Oil Red O dye solution for 30 min, washed with deionized water (ddH2O) five times, and then photographed under a microscope. Next, 200 μL of isopropyl alcohol was added to each well; the cells oscillated on a shaker for 5 min. Finally, 200 μL of liquid was sucked into a 96-well plate and read at a 510 nm wavelength for the quantitative analysis.
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