Description
The dialdehydes formed by treatment with periodic acid from certain carbohydrates, will selectively reduce an alkaline hexamine-silver salt mixture. This kit is especially suitable for renal glomerular basement membranes. It is very helpful to determine the thickness of glomerular basement membranes and mesangial matrix.
Kit Components
| Sku# | Reagent Name | Volume (ml) |
| VB-3033-1 | Periodic acid 1% | 100 ml |
| VB-3033-2 | Silver nitrate 5% | 35 ml |
| VB-3033-3 | Methenamine 3% | 35 ml |
| VB-3033-4 | Sodium tetraborate 2% | 35 ml |
| VB-3033-5 | Gold chloride 0.25% | 100 ml |
| VB-3033-6 | Sodium thiosulfate 5% | 100ml |
| VB-3033-7 | Mayer’s hematoxylin solution | 100 ml |
| VB-3033-8 | Eosin solution | 100 ml |
Storage
Store Silver Nitrate 5% and Gold chloride 0.25% at 2-8 °C. The other components can be stored at room temperature.
Protocol
- Deparaffinize in xylene I for 6 minutes and II for 6 minutes. Rehydrate:
- Ethanol 100% (2 minutes)×2
- Ethanol 95% (2 minutes)×2
- Ethanol 70% (2 minutes)×1
- Rinse in DI water (2 minutes)×1
- Incubate the sections with Periodic acid 1% for 30 minutes.
- Wash in distilled water.
- Fresh make silver methenamine solution: Mix Silver nitrate 5% , Methenamine 3% and Sodium tetraborate 2% at a ratio of 1:1:1.
- Incubate the section with silver methenamine solution in oven for 30 minutes at 60°C. Check the slide microscopically after 20 minutes then every 5 minutes until the basement membrane is well demonstrated. If necessary, incubate again. The sections should turn tobacco.
- Wash in hot distilled water (60°C) for 4 min. Wash in room temperature distilled water for 1 min.
- Incubate the section with Gold chloride 0.25% for 1 minute.
- Wash in distilled water.
- Incubate the section with Sodium thiosulfate 5% for 1 minute.
- Wash in distilled water.
- Stain the sections with hematoxylin for 2 min.
- Running tap water for 2 min.
- Stain the sections with Eosin for 1min.
- Dehydrate with 2 changes of 100% Ethanol (2 minute per change).
- Clear with 2 changes of xylene (5 minute per change).
- Mount cover slip onto glass slide with Permount or some other suitable organic mounting medium.
Control Tissue: Kidney section
Expected Results:
- Basement membranes …………………………….Black
- Cell Nuclear…………………………………………Blue
- Background ………………………………………..Pink/Red
References:
- Ohga S, et al (2007) . Thiazolidinedione ameliorates renal injury in experimental diabetic rats through anti-inflammatory effects mediated by inhibition of NF-kappaB activation. American journal of physiology. Renal physiology 292(4):F1141-50.
- Francis A, at al.(2016). Polypoid Change of the Glomerular Basement Membrane in a Child with Steroid Resistant Nephrotic Syndrome and ARHGAP24 Mutation: A Case Report. The Open Urology & Nephrology Journal 9(1):88-93
Note:
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Precautions:
Handle with care. Avoid contact with eyes, skin and clothing. Do not ingest. Wear gloves.
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