Copper Blood Staining Kit
Cat No.: DCP-CBSK
Size: 50 tests (compatible with 4 μm-thick sections)
Ultrapure, Filtered 0.1-micron membrane TWICE and 0.04-micron membrane ONCE.
Description:
The Copper Histological Staining Kit is a precision-engineered solution designed for the reliable detection of copper deposits in formalin-fixed, paraffin-embedded tissue sections. Optimized for diagnostic applications such as Wilson’s disease identification and research into copper metabolism disorders, this kit combines industry-standard protocols with enhanced stability for consistent, high-quality results. The kit streamlines the staining process into a 90-minute protocol, including deparaffinization, hydration, rhodanine incubation, and counterstaining. The microwave-optimized variant completes staining in under 15 minutes, ideal for high-throughput labs. All solutions are compatible with automated stainers, and the xylene-free dehydration series reduces occupational hazards.
Applications:
1. Electrophoresis and Western Blotting
- Diagnostic Pathology: Gold-standard detection of hepatic copper in Wilson’s disease, primary biliary cholangitis, and copper toxicosis.
- Research: Study copper distribution in neurodegenerative models (Alzheimer’s, Parkinson’s) and oncology (copper dysregulation in tumors).
- Veterinary Medicine: Identify copper storage disorders in canine and ovine specimens.
2. Forensic Bloodstain Enhancement
- Latent Blood Visualization: Reacts with hemoglobin and serum albumin on porous/non-porous surfaces, producing permanent blue-black patterns. Compatible with DNA preservation protocols, unlike luminol-based methods.
- Reduced Background: Acetic acid minimizes nonspecific binding on semi-porous materials (e.g., wood, fabric), improving contrast compared to methanol blends.
3. Histological Staining
- Collagen Specificity: Binds selectively to collagen fibers in tissue sections, enabling differential visualization of connective tissues in 10–15 minute protocols.
Advantages:
- High Sensitivity: Detects copper concentrations as low as 0.1–0.5 μg/g tissue, ensuring accurate identification of early pathological copper accumulation.
- Rhodanine-Based Staining: Utilizes a 0.2% alcoholic rhodanine solution for selective chelation of free and protein-bound copper, producing stable red-brown complexes under standard microscopy.
- Rapid Protocols: Includes both conventional (2-hour heated incubation) and accelerated microwave-assisted (6-minute) workflows to accommodate urgent diagnostic needs.
- Integrated Controls: Pre-stained fetal liver sections (50–100 μg Cu/g) and preparation guidelines for spiked positive controls ensure batch-to-batch reliability.
- Ready-to-Use Components: Pre-mixed acetate buffer (pH 8.0) and oxidized Mayer’s hematoxylin reduce preparation time while maintaining pH stability.
Kit Components:
| Solution |
Size |
| Rhodanine Solution |
30 mL |
| Acetate Buffer |
500 mL |
| Mayer’s Hematoxylin |
125 mL |
Additional materials required (not included):
-
Xylene, ethanol series (100%, 95%, 70%), distilled water, Coplin jars, microwave (optional), mounting medium.
Protocol:
A. Standard Method (2-Hour Incubation)
-
Deparaffinization
-
Xylene I: 6 min
-
Xylene II: 6 min
-
Hydration
-
100% ethanol: 2 min ×2
-
95% ethanol: 2 min ×2
-
70% ethanol: 2 min
-
Distilled water: 2 min
-
Rhodanine Staining
-
Rinsing
-
Counterstaining
-
Differentiation
-
Dehydration & Mounting
-
70% ethanol: 2 min
-
95% ethanol: 2 min ×2
-
100% ethanol: 2 min ×2
-
Xylene: 5 min ×2
-
Mount with resin medium.
B. Accelerated Microwave Protocol
-
Deparaffinize/Hydrate (as above).
-
Microwave Staining
-
Heat 100 mL water in microwave-safe jar to 90°C (1–2 min at 700W).
-
Place slide over steam, apply 5 drops Working Rhodanine Solution.
-
Microwave at 700W in 30-sec cycles (3–4 cycles total). Cool between cycles.
-
Rinse, Counterstain, & Mount (follow Steps 4–7 above).
Quality Control:
- Sterility: Filtered 0.1 micron Twice and 0.04 micron once.
- DNase activity: None was detected after incubation of plasmid DNA, and this product was used for 18 hours at room temperature.
- RNase activity: No RNase activity was detected after incubation of ribosomal RNA and this product for 18 hr at room temperature.
Storage and Shelf Life:
| Component |
Condition |
Shelf Life |
| Rhodanine Solution |
2–8°C (amber glass) |
6 months |
| Acetate Buffer |
15–25°C |
12 months |
| Hematoxylin |
15–25°C |
18 months |
Disposal:
CUSTOMIZATION: Please INQUIRE if other concentrations, additions of chemicals/compounds/proteins/supplements, different pH, and modifications are needed.
All DiagnoCine Precision Sterile buffers are filtered-sterilized with a 0.1-micron filtration two times and 0.04-micron filtration one time. Thus, mycoplasma contamination is prevented. The smallest size mycoplasma type can be about 0.2 microns.
* This product is manufactured under ISO 13485-certified and CE-approved facilities (Suppliers of DiagnoCine Precision). All final packaging, quality assurance, and testing are done at the DiagnoCine R&D and Quality Testing Center. All specific customization requests and assembly were accomplished at DiagnoCine Precision in Totowa, New Jersey, USA.
