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| Product Name | Sudan Black B Lipid Stain Solution, for Leukocyte Granules and Lipids |
| Catalog No. | ATRSA-0029 |
| Description | Ready-to-use Sudan Black B staining solution for histochemical detection of intracellular lipids, phospholipids, and lipoproteins in hematological and tissue specimens. Sudan Black B is a slightly basic lysochrome (lipid-soluble dye) that partitions from solvent into lipid-rich structures. In hematology, it principally stains primary (azurophilic) and secondary (specific) granules in granulocytes and their precursors, weakly stains monocyte granules, and does not stain lymphoid cells. This staining pattern aids in the classification of acute leukemias (distinguishes AML with granulocytic differentiation from ALL) and identification of cells of granulocytic lineage in myelodysplastic syndromes and myeloproliferative neoplasms. |
| Intended Use | Histochemical staining of lipids in blood and bone marrow smears for hematological malignancy classification; detection of lipid droplets in tissue sections for steatosis assessment and adipose tissue analysis; bacterial lipid inclusions (PHB granules) staining. |
| Principle / Technology | Sudan Black B (C29H24N6) is a lysochrome that dissolves in and stains neutral lipids, phospholipids, and sterols; staining mechanism is primarily physical (partitioning/solubility) rather than chemical; counterstaining with nuclear fast red or Giemsa provides morphological contrast. |
| Detection Method | Fix air-dried blood/bone marrow smear in formalin vapor or 37% formaldehyde (10 min); rinse; stain in Sudan Black B working solution (30-60 min at RT); rinse in 70% ethanol briefly to differentiate; wash in water; counterstain with nuclear fast red or Giemsa (5-10 min); wash, dry, mount. |
| Sample Type | Peripheral blood smears, bone marrow aspirate smears, frozen tissue sections (10-20 µm), cytospin preparations. |
| Performance Range / Specifications | Staining pattern: granulocytes — heavy black granular staining; monocytes — fine scattered black granules or negative; lymphocytes — negative; eosinophil granules stain brown-black; basophil granules may be negative or weakly positive. |
| Sensitivity / LOD | Detection of as few as 1-5% Sudan Black B-positive blasts in a leukemic population; lipids detectable at microgram levels in tissue sections. |
| Specificity | Stains phospholipids, neutral lipids (triglycerides), and sterols; does not distinguish between lipid classes; formalin fixation may reduce phospholipid staining but does not affect neutral lipid staining; weak or negative staining of cholesterol esters. |
| Reaction Conditions / Protocol | Stain at room temperature; time-dependent intensity (30 min minimum, up to overnight for maximum sensitivity); controlled differentiation in 70% ethanol removes excess dye from non-lipid structures. |
| Components / Formulation | Sudan Black B stock solution (saturated in 70% ethanol or propylene glycol), Sudan Black B working solution (filtered), nuclear fast red counterstain (0.1%), mounting medium (aqueous-based). |
| Storage Conditions | Store stock solution at room temperature in tightly capped amber bottle; working solution prepare fresh or use within 2 weeks. |
| Shelf Life | 12 months from date of manufacture for stock solution. |
| Package Specifications | 100 mL stock solution (sufficient for ~200 slide preparations), 500 mL bulk size. |
| Product Form | Stock solution: dark blue-black liquid with visible dye particles (filter before use); counterstain: pink-red liquid. |
| Quality Control | Each lot tested on normal peripheral blood smear (neutrophil granules intensely black, lymphocytes negative) and on AML M2 bone marrow smear (Sudan Black B reactivity of blasts). |
| Key Features | Ready-to-use stock solution; classic lipid staining for hematological classification; distinguishes AML from ALL; stable at room temperature; nuclear fast red counterstain included; aqueous mounting medium — no xylene required. |
| Purity | Sudan Black B dye content per specification; filtered working solution free of particulate dye aggregates. |
| Concentration | Saturated solution in 70% ethanol; typical working concentration ~0.5-1% w/v. |
| Activity / Unit Definition | Lipid staining capacity verified by positive reaction with oleic acid and lecithin spots on filter paper. |
| Molecular Weight | Sudan Black B: 456.54 g/mol (C29H24N6). |
| Source / Origin | Synthetic diazo dye; counterstain from natural hematoxylin derivative. |
| pH Range / Optimal pH | Staining solution neutral pH (70% ethanol based). |
| Shipping Conditions | Ambient temperature; protect from light. |
| Expiration Date / Stability | 12 months for stock solution at RT; working solution use within 2 weeks (filter before each use). |
| Regulatory / Compliance | For in vitro diagnostic use in hematology and histology; manufactured under ISO 13485 quality system. |
| Compatibility | Compatible with air-dried blood and bone marrow smears. Formalin vapor fixation preferred over immersion. Not recommended for paraffin-embedded sections (paraffin dissolves lipids during processing) — use frozen sections. Can be combined with myeloperoxidase (MPO) and nonspecific esterase (NSE) for comprehensive AML classification panel. |
| Recommended Buffer System | Not buffered; 70% ethanol-based dye solution. |
| Application Notes / Precautions | Always filter working solution through Whatman #1 filter paper before use to remove dye precipitates that cause artifact deposits on slides. Fresh smears (within 1 week) give best results. Over-differentiation in ethanol removes specific staining — monitor under microscope. Sudan Black B staining of normal granulocytes serves as an internal positive control. For cytochemistry panel, perform MPO, Sudan Black B, and NSE on separate slides from the same sample. Mount slides with aqueous mounting medium (glycerin jelly or aqueous permanent mountant). |
| Batch-to-Batch Consistency | Dye solubility and staining intensity within specification using standardized leukocyte preparation; Sudan Black B content verified by spectrophotometry. |
For research use only, not for clinical use.
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