The cells were fixed for 30 min at room temperature in 4% paraformaldehyde. Subsequently, Oil Red O (Servicebio, Wuhan, China) was mixed with deionized water (3:2) and then added to the cells for 15 min. Ultimately, PBS was used to clean the aforementioned cells until no visible impurities remained. Images were captured using an inverted microscope (Olympus, Tokyo, Japan).
Oil red o
Manufactured by Wuhan Servicebio Technology
Sourced in China
Oil Red O is a fat-soluble dye commonly used in histological staining techniques. It selectively stains neutral lipids, such as triglycerides and cholesterol esters, making it a useful tool for the identification and visualization of lipid-rich structures in cells and tissues.
Automatically generated - may contain errors
Lab products found in correlation
Oct compound, by Merck Group (3 mentions)
Eos 7d digital camera, by Canon (3 mentions)
Cryostar nx50, by Thermo Fisher Scientific (2 mentions)
Inverted microscope, by Olympus (1 mentions)
Tg assay kit, by Applygen (1 mentions)
Dp71 microscope, by Olympus (1 mentions)
Glycerol jelly mounting medium, by Beyotime (1 mentions)
Hematoxylin, by Beyotime (1 mentions)
Tissue tek optimal cutting temperature compound, by Sakura Finetek (1 mentions)
Sirius red, by Wuhan Servicebio Technology (1 mentions)
Maxvision hrp polymer anti mouse rabbit ihc kit, by Maixin Group (1 mentions)
Periodic acid schiff staining, by Solarbio (1 mentions)
Hematoxylin and eosin h e staining, by Solarbio (1 mentions)
Proteinase k solution, by Solarbio (1 mentions)
Tunel reaction mixture, by Roche (1 mentions)
Fluorescence microscope, by Nikon (1 mentions)
Isopropyl alcohol, by Merck Group (1 mentions)
G1101, by Wuhan Servicebio Technology (1 mentions)
Light microscopy, by Olympus (1 mentions)
Dm500b, by Leica (1 mentions)
34 protocols using oil red o
1
Oil Red O Staining of Lipids
2
Quantitative Adipocyte Lipid Analysis
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3T3-L1 adipocytes and preadipocytes were stained with oil red O (Servicebio) for 15 min and washed with PBS. The positively stained cells were captured under the microscope. To measure the lipid accumulation, oil red O stained cells were washed with isopropyl alcohol and supernatants were tested by Multiscan Spectrum at 492 nm.
3
Histological Quantification of Hepatic Lipid Accumulation
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The fixed liver tissues were stained with Oil Red O (Servicebio, Wuhan, China) for fat coloring. The slide was rinsed with 75% of ethyl alcohol for histo-differentiation and then washed with distilled water. The nuclei of the stained tissues were dyed blue with hematoxylin. Sections were examined with the NIKON ECLIPSE TI-SR inverted fluorescence microscope (NIKON, Tokyo, Japan), and scanned sections were captured using the NIKON DS-U3 microscope camera controller (NIKON, Tokyo, Japan). Mean adipose area was quantified with 3 tissue sections for each animal using Image J (v1.8.0).
4
Liver and cell triglyceride detection
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Liver and cell TG content was detected by a TG assay kit (Applygen Technology, Inc., Beijing, China) in accordance with the manufacturer's instructions. Ten microliters of liver frozen sections were stained with Oil Red O (Servicebio, Wuhan, China). The slides were viewed at 200× and 400× magnification.
5
Quantitative Oil Red O Staining for Lipid Assessment
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Oil Red O (ORO) was performed conventionally. Briefly, frozen PTC and para-cancer tissues were cryosectioned at 10 μm thickness on a freezing microtome (CRYOSTAR NX50, Thermo, China) and stained with Oil Red O(Servicebio, China) for 10 min. After rinsing in distilled water, sections were counterstained for 5 min in hematoxylin. The percentage of ORO-stained cells was determined (0, 0–5%; 1, 5–25%; 2, 25–50%; 3, 50–75%; 4, >75%).
6
Frozen Placenta Analysis using Oil Red O
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The frozen placenta was embedded in Tissue-Tek O.C.T.Compound. Samples were cut at a thickness of 10 μm from each sample. Slides were washed in distilled water for 2 min and 60% isopropanol for 2 min. Afterward, slides were stained for 10 min in 60% working Oil Red O (G1015, ServiceBio, Beijing, China). Then slides were washed shortly with distilled water and 60% isopropanol, stained for 1 min with Hematoxylin (C0107, Beyotime, Shanghai, China), washed in running water for 10 min, mounted using glycerol jelly mounting medium (C0187, Beyotime, Shanghai, China). Images were captured using an Olympus DP71 microscope. Five random fields of view per tissue section were used to quantify the integrated optical density (IOD) using ImageJ software.
7
Comprehensive Liver Tissue Analysis
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The liver was embedded with tissue-Tek optimal cutting temperature compound (Sakura Finetek, Japan), or paraffin, and liver sections were prepared. Liver sections were used for H&E, Sirius Red, and Oil Red O staining (Servicebio, Wuhan, China). For immunohistochemistry, antigen retrieval was performed under high pressure and temperature in 0.01 mmol/L citrate buffer (pH = 6.0). Then, the sections were then incubated with 3% H2O2 at room temperature for 10 min to block the endogenous peroxidase activity. After blocked with 1% BSA, the sections were incubated with primary antibodies overnight at 4 °C and then secondary antibodies were used for 1 h at 37 °C. Immunochemical staining of collagen type I (Col1α), CD68, and 4 hydroxynonenal (4-HNE) was performed using paraffin-embedded liver sections. Detailed information about antibodies is listed in Supporting Information Table S3 . Quantitative analysis was performed using open-source software (Image-J software (http://imagej.nib.gov/ )).
8
Comprehensive Kidney Tissue Analysis
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The kidney tissues underwent dehydration, paraffin embedding, and were cut into 3 μm thick sections using a microtome. Morphological changes in the kidney tissues were observed through periodic acid-Schiff (PAS) staining (Solarbio, Beijing, China) and hematoxylin and eosin (H&E) staining (Solarbio, Beijing, China). The mitochondrial ultrastructure of the kidney tissue was examined using transmission electron microscopy (TEM).
Following sectioning, the kidney tissues were sliced into 5 μm sections and stained with Oil Red O (Servicebio, Wuhan, China) to assess the oil content in the kidney tissue. For immunohistochemical (IHC) studies, paraffin-embedded kidney sections were dewaxed, rehydrated, blocked, and incubated overnight with PGC-1α antibody (Invitrogen, CA, USA). The sections were then incubated with the MaxVision TM HRP-Polymer anti-Mouse/Rabbit IHC Kit (Maixin, Fuzhou, China), stained with diaminobenzidine (Solarbio, Beijing, China), sealed, and observed under a microscope. Kidney sections intended for TUNEL staining were sequentially immersed in xylene and alcohol, followed by incubation in Proteinase K solution (Solarbio, Beijing, China). TUNEL reaction mixture (ROCHE, Basel, Switzerland) was added to the sections, incubated in a dark chamber, and subsequently examined for apoptosis in the kidney sections.
Following sectioning, the kidney tissues were sliced into 5 μm sections and stained with Oil Red O (Servicebio, Wuhan, China) to assess the oil content in the kidney tissue. For immunohistochemical (IHC) studies, paraffin-embedded kidney sections were dewaxed, rehydrated, blocked, and incubated overnight with PGC-1α antibody (Invitrogen, CA, USA). The sections were then incubated with the MaxVision TM HRP-Polymer anti-Mouse/Rabbit IHC Kit (Maixin, Fuzhou, China), stained with diaminobenzidine (Solarbio, Beijing, China), sealed, and observed under a microscope. Kidney sections intended for TUNEL staining were sequentially immersed in xylene and alcohol, followed by incubation in Proteinase K solution (Solarbio, Beijing, China). TUNEL reaction mixture (ROCHE, Basel, Switzerland) was added to the sections, incubated in a dark chamber, and subsequently examined for apoptosis in the kidney sections.
9
Aortic Lipid Accumulation Quantification
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Lipid accumulation of the thoracoabdominal aorta was determined by oil red O staining (Servicebio, G1015). Aorta was fixed with 4% paraformaldehyde for at least 24 h, then washed with PBS. After soaking 60% isopropyl alcohol for 3 s, the aorta was stained by oil red O at 37°C for 30 min. The excess oil red O was removed by using 60% isopropyl alcohol. Lastly, the samples with a scale were photographed and quantified.
10
Visualizing Lipid Droplets in Liver Tissue
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To visualize lipid droplets in the liver, frozen liver tissues from anesthetized mice were embedded in Tissue-Tek OCT compound, and then sliced to 8–10 μm thick frozen sections. Frozen sections were fixed with fixative (G1101, servicebio, Wuhan, China) and stained with Oil Red O (G1016, servicebio, Wuhan, China) without light for 8–10 min. Then the tissue sections were washed with 60% isopropyl alcohol (#I9030, Sigma-Aldrich) and re-stained with hematoxylin. The slides were sealed with glycerin-glutin. For cell ORO staining, cells were wash by PBS (SH30256.01, HyClone, United States) and fixed with 4% paraformaldehyde, then cells were stained with ORO as described above. Images were taken with the Nikon Fluorescence Microscope.
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