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| Product Name | Luxol Fast Blue Myelin Stain Kit, Modified Kluver-Barrera Method |
| Catalog No. | ATRSA-0030 |
| Description | Myelin staining kit based on the Luxol Fast Blue (LFB) technique with cresyl violet counterstain (modified Kluver-Barrera method) for selective visualization of myelinated nerve fibers in brain and spinal cord tissue sections. LFB, a copper phthalocyanine dye, binds to phospholipid components of myelin through ionic and hydrophobic interactions, producing an intense blue to turquoise staining of myelinated fibers with excellent contrast against grey matter. The cresyl violet counterstain simultaneously labels Nissl substance (rough endoplasmic reticulum) in neuronal cell bodies and glial nuclei in shades of violet, enabling concurrent assessment of myelin integrity and neuronal morphology. |
| Intended Use | Histological visualization of myelin distribution and integrity in brain and spinal cord paraffin or frozen sections for neuropathology studies, demyelinating disease models (multiple sclerosis, leukodystrophy), neurotoxicity assessment, and developmental neurobiology. |
| Principle / Technology | Luxol Fast Blue is a copper phthalocyanine dye that binds phospholipid components of myelin sheaths; differentiation in lithium carbonate solution removes excess dye from grey matter while myelin retains stain due to high phospholipid content; cresyl violet counterstain labels Nissl substance (RNA in RER) in neuronal cytoplasm violet-blue. |
| Detection Method | Deparaffinize and hydrate sections; stain in Luxol Fast Blue solution at 56-60 °C overnight (16-24 hours); cool to RT; rinse in 95% ethanol and distilled water; differentiate in 0.05% lithium carbonate (5-10 sec) followed by 70% ethanol until grey-white matter distinction; wash; counterstain with cresyl violet (5-10 min at 60 °C); dehydrate, clear, mount. |
| Sample Type | Formalin-fixed paraffin-embedded (FFPE) brain and spinal cord sections (5-15 µm); frozen sections (20-40 µm) with modified protocol. |
| Performance Range / Specifications | Staining result: myelinated white matter — intense blue to turquoise; grey matter — pale blue to unstained; neuronal Nissl substance — violet; glial nuclei — violet; demyelinated lesions — pale or unstained (loss of blue). |
| Sensitivity / LOD | Detectable myelin in fine fibers of cortical and deep grey matter structures; capable of visualizing early demyelination before axonal loss. |
| Specificity | LFB specifically stains myelin phospholipids; high specificity for CNS myelin; also stains PNS myelin; does not stain non-myelinated axons; cresyl violet specificity for RNA-rich Nissl substance enables neuronal morphology assessment. |
| Reaction Conditions / Protocol | LFB staining: overnight at 56-60 °C; differentiation: lithium carbonate (5-10 sec) / 70% ethanol; cresyl violet: 5-10 min at 60 °C; complete protocol including embedding takes ~24 hours. |
| Components / Formulation | Luxol Fast Blue MBS solution (0.1% in 95% ethanol with 0.5% glacial acetic acid), lithium carbonate solution (0.05%), cresyl violet acetate solution (0.1%, pH 3.8-4.0), detailed protocol. |
| Storage Conditions | Store LFB solution at room temperature in tightly capped amber bottle; cresyl violet at room temperature; lithium carbonate at room temperature. |
| Shelf Life | 12 months from date of manufacture. |
| Package Specifications | 1 kit (sufficient for ~100 slides), 5 kits. |
| Product Form | LFB: dark blue alcoholic solution; lithium carbonate: clear liquid; cresyl violet: purple aqueous solution. |
| Quality Control | Each lot tested on standardized mouse/rat brain coronal sections for myelin staining intensity, grey-white matter differentiation, and cresyl violet counterstaining of hippocampal pyramidal neurons. |
| Key Features | Classic Kluver-Barrera method; simultaneous myelin + Nissl staining; copper phthalocyanine dye for high specificity; excellent grey-white matter contrast; compatible with paraffin and frozen sections; 24-hour protocol; permanent mountable preparation. |
| Purity | Luxol Fast Blue MBS dye content verified; cresyl violet acetate certified for histology; lithium carbonate ACS grade. |
| Concentration | LFB: 0.1% w/v in acidified 95% ethanol; cresyl violet: 0.1% w/v in acetate buffer pH 3.8. |
| Activity / Unit Definition | Myelin staining verified by densitometric analysis of corpus callosum vs. cortex. |
| Molecular Weight | Luxol Fast Blue MBS: varies (copper phthalocyanine sulfonated derivative). |
| Source / Origin | Synthetic copper phthalocyanine dye; cresyl violet from synthetic organic synthesis. |
| pH Range / Optimal pH | LFB solution acidified with glacial acetic acid; cresyl violet critical pH 3.8-4.0 for Nissl specificity; lithium carbonate alkaline pH ~11. |
| Shipping Conditions | Ambient temperature; protect from light. |
| Expiration Date / Stability | 12 months at RT; cresyl violet may develop precipitate over time — filter before use. |
| Regulatory / Compliance | For research and histopathological use; not for diagnostic procedures without appropriate pathology laboratory validation. |
| Compatibility | Compatible with formalin-fixed paraffin-embedded tissue sections (5-10 µm for rodent, 5-15 µm for human). For frozen sections: reduce staining time to 2-4 hours at 56 °C. Best results on perfusion-fixed tissue. Not recommended for tissues fixed in Bouin's or other acidic fixatives. |
| Recommended Buffer System | Cresyl violet uses sodium acetate buffer, pH 3.8-4.0. |
| Application Notes / Precautions | LFB solution must be preheated to 56-60 °C and sections must be completely deparaffinized. Differentiation is the most critical step — lithium carbonate treatment should be 5-10 seconds only; check microscopically and repeat if needed. After differentiation, immediately rinse in 70% ethanol to stop the reaction. For demyelination models, ensure sham control sections are processed in parallel for comparison. Cresyl violet working solution should be filtered before use and can be reused for ~10 staining runs. |
| Batch-to-Batch Consistency | LFB dye concentration within specification; cresyl violet pH within ±0.1 units; staining performance verified on reference brain sections per lot. |
For research use only, not for clinical use.
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