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| Product Name | Oil Red O Stain Kit for Frozen Sections and Cultured Cells |
| Catalog No. | ATRSA-0031 |
| Description | Histochemical staining kit for detection and visualization of neutral lipids, triglycerides, and cholesteryl esters in frozen tissue sections and cultured cells using the Oil Red O (ORO) lysochrome dye. Oil Red O is a fat-soluble diazo dye that partitions from its alcoholic solvent into lipid droplets more readily than into the surrounding aqueous environment, resulting in intense red staining of lipid-rich structures. The kit is optimized for frozen sections (paraffin processing removes lipids) and cultured adipocytes/hepatocytes, providing an essential tool for steatosis quantification, adipogenesis assessment, atherosclerosis plaque analysis, and lipid metabolism research. |
| Intended Use | Histological detection and semi-quantitative analysis of neutral lipid accumulation in frozen liver sections for steatosis grading, frozen arterial sections for atherosclerosis analysis, cultured adipocytes for adipogenesis assessment, and cultured hepatocytes for lipotoxicity studies. |
| Principle / Technology | Oil Red O is more soluble in neutral lipids than in its alcoholic/propylene glycol solvent system; dye partitions into lipid droplets producing intense red color; hematoxylin counterstain provides nuclear contrast; staining is primarily physical (partitioning) rather than chemical. |
| Detection Method | Prepare frozen sections (8-15 µm), air dry; fix in 10% formalin or 4% PFA (10 min); rinse with water; rinse with 60% isopropanol (optional, for propylene glycol method); stain in pre-warmed ORO working solution (10-15 min at RT or 60 °C); rinse in 60% isopropanol (differentiate); wash with water; counterstain with hematoxylin (1-3 min); wash, mount with aqueous/glycerin mounting medium. |
| Sample Type | Fresh frozen tissue sections (liver, aorta, muscle, adipose), cultured cells grown on coverslips or chamber slides, isolated lipid extracts on TLC plates. |
| Performance Range / Specifications | Staining result: neutral lipid droplets — intense red to orange-red; nuclei — blue/purple (hematoxylin counterstain); cytoplasm — unstained to pale pink; detection from microvesicular (small droplet) to macrovesicular (large droplet displacing nucleus) steatosis. |
| Sensitivity / LOD | Detectable lipid droplets as small as 0.5 µm diameter; microvesicular steatosis visible at 200-400× magnification; macroscopically visible steatosis >30% parenchymal involvement. |
| Specificity | Oil Red O stains neutral lipids and cholesteryl esters (deep red) and some phospholipids (pale pink/orange); does not stain free cholesterol or fatty acids without prior conversion; specificity enhanced by lipid extraction control slide (acetone or chloroform-methanol pre-treatment). |
| Reaction Conditions / Protocol | Fix sections 10 min at RT; stain 10-15 min at RT; differentiate in 60% isopropanol (5-15 sec); counterstain 1-3 min; complete in <1 hour. |
| Components / Formulation | Oil Red O stock solution (0.5% in isopropanol or 0.3% in propylene glycol), hematoxylin counterstain (Mayer's or Gill's), 60% isopropanol for differentiation, aqueous mounting medium (glycerin jelly). |
| Storage Conditions | Store ORO stock at room temperature protected from light; hematoxylin at room temperature; mounting medium at 2-8 °C. |
| Shelf Life | 12 months from date of manufacture. |
| Package Specifications | 1 kit (sufficient for ~200 frozen sections), 5 kits. |
| Product Form | ORO stock: dark red liquid; hematoxylin: dark purple liquid; aqueous mounting medium: viscous liquid. |
| Quality Control | Each lot tested on standardized mouse liver frozen section (high-fat diet) for lipid droplet staining intensity and hematoxylin nuclear counterstain contrast. |
| Key Features | Classic lipid detection with Oil Red O; optimized for frozen sections; hematoxylin counterstain included; aqueous mounting medium (no xylene); room temperature protocol; suitable for liver, artery, and cell culture; semi-quantitative steatosis grading compatible. |
| Purity | Oil Red O dye content ≥75% (dye basis); hematoxylin certified for histology. |
| Concentration | Oil Red O stock: 0.5% w/v in isopropanol or 0.3% w/v in propylene glycol; working solution: 3 parts stock + 2 parts water, filtered. |
| Activity / Unit Definition | Lipid staining: binding capacity verified by filter paper spot test with triolein standard. |
| Molecular Weight | Oil Red O: 408.49 g/mol (C26H24N4O). |
| Source / Origin | Synthetic monoazo dye; hematoxylin from logwood (Haematoxylum campechianum) extract. |
| pH Range / Optimal pH | ORO solution neutral; hematoxylin pH ~2.5 (acidic). |
| Shipping Conditions | Ambient temperature. |
| Expiration Date / Stability | 12 months at RT; ORO working solution prepare fresh and use within 2 hours (dye precipitates over time). |
| Regulatory / Compliance | For research and histopathological use; manufactured under ISO 9001 quality system. |
| Compatibility | Optimized for fresh frozen tissue sections cut at 8-15 µm thickness. Not compatible with paraffin-embedded sections (paraffin removes lipids). Compatible with cells cultured on glass coverslips, chamber slides, or plastic culture dishes (for plastic, use lower isopropanol concentration for differentiation to avoid etching). Can be combined with immunohistochemistry if ORO staining is performed before IHC. |
| Recommended Buffer System | Not applicable — alcoholic dye solution. |
| Application Notes / Precautions | Always prepare ORO working solution fresh and filter through Whatman #1 before use — dye precipitates cause artifact. Cut frozen sections fresh and air dry thoroughly before fixation. Over-differentiation in 60% isopropanol will remove specific lipid staining — dip quickly (1-5 seconds). Mount immediately with aqueous mounting medium as ORO fades in organic mounting media. Include a lipid extraction control slide (acetone treatment, 5 min) to confirm staining specificity. For quantitative steatosis analysis, use ImageJ or similar software on captured images. |
| Batch-to-Batch Consistency | ORO dye solubility and staining intensity within specification; hematoxylin nuclear staining intensity within expected range per lot. |
For research use only, not for clinical use.
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