Campbell-Switzer Alzheimer Silver Stain
Plaques and Tangles: A Controlled and Enhanced Method
Staining Procedure:
(125×65 dishes for up to 30 sxns max & 100×50 dishes for 13 or less sxns.)
- Prepare Physical Developer Solutions A, B & C
- Nitric acid wash all glassware.
- Remove sections from storage containers and place in dH2O 3 x 10 min each. Use baskets to transfer sections through all steps except the SPC and Physical Developer ABC steps.
- Prepare SPC Solutions. Stir until needed. Prepare other solutions.
- Place sections in 2% NH4OH. Stir on stirring table for 5 min.
- Place in dH2O 2 x 1 min. each.
- Place in the SPC Soln. and cover. Stir gently for 40 min.
- Place in 1% Citric Acid for 3 min.
- Place in 4.99 pH Acetate Buffer Working Solution until ready for Physical Developer.
- Prepare fresh Physical Developer ABC Sol’n. for each batch. Develop 10-15 sections at a time. Use fewer sections if they appear crowded.
- Place in Physical Developer ABC Sol’n. over a light source. The development time is visually assessed.
- Stop development by placing in 4.99 pH Acetate Buffer Working Solution briefly.
- Place in fresh 4.99 pH Acetate Buffer Working Solution. Timing is not critical.
- Place in fresh 4.99 pH Acetate Buffer Working Solution. Timing is not critical.
- Place into fresh dH2O 30 sec.
- Place into 0.5% Sodium Thiosulfate solution for 45 sec.
- Place into fresh dH2O 3 x 2 min. each
- Mount sections.
- Coverslip.
Other links of possible interest:
- The original abstract introducing the Campbell-Switzer method quantitative analysis of staining results
- About the Campbell-Switzer method
- For comparison of CS stain with antibody stains See Schonheit et al
- NSA offers digital, systematic analysis of the stained sections from the Campbell-Switzer stain. For details, refer to our Image Analysis page.
FAQ regarding silver stains:
No. There are numerous silver staining protocols, each tailored to reveal specific features. "Silver Stains" are as unique as any other stain in that they are each specifically designed to stain a particular feature. Silver simply refers to the staining agent (Silver) which stains the designated feature black. See: Switzer, R. C. III. Application of Silver Degeneration Stains to Neurotoxicity Testing. Toxicologic Pathology 28: 70-83, 2000.
No. Each method reveals a different set of features. The Campbell-Switzer method reveals the neuritic plaques and tangles of Alzheimer's pathology. The disintegrative degeneration stain shows the products of neuronal disintegrative degeneration.
Some antibodies show only a subset of plaques revealed by the Campbell-Switzer method, while others appear to be a one to one match. The Campbell-Switzer stain is unique in its ability to differentiate its staining of immature (diffuse) and mature (congophilic) plaques, allowing researchers a unique observational capability.
For further reference, please see: Stains
Solutions:
| Silver Stain Solution #1: 1% Silver Nitrate (AgNO3) | |
|---|---|
| 100 ml dH2O | Prepare and mix well. |
| 1.0 g AgNO3 | |
| 100 ml 1% AgNO3 |
| Silver Stain Solution #2: 1% Potassium Carbonate (K2CO3) | |
|---|---|
| 100 ml dH2O | Prepare and mix well. |
| 1.0 g K2CO3 | |
| 100 ml 1% K2CO3 |
| Silver Stain Solution #3: Silver-Pyridine-Carbonate (SPC) Batch Sizes | ||||||
|---|---|---|---|---|---|---|
| 1% AgNO3 | 60 ml | 120 ml | 180 ml | 240 ml | 300 ml | 360 ml |
| Pyridine | 17 ml | 34 ml | 51 ml | 68 ml | 85 ml | 102 ml |
| 1% K2CO3 | 45 ml | 90 ml | 135 ml | 180 ml | 225 ml | 270 ml |
| Total Volume: | 122 ml | 244 ml | 366 ml | 488 ml | 610 ml | 732 ml |
| Silver Stain Solution #4: 2% Ammonium Hydroxide | |
|---|---|
| 98 ml dH2O | Prepare and mix well. |
| 2 ml NH4OH (27-29% concentrate) | |
| 100 ml 2% NH4OH |
| Silver Stain Solution #5: 1% Citric Acid | |
|---|---|
| 100 ml dH20 | Prepare and mix well. |
| 1.0 g Citric Acid | |
| 100 ml 1% Citric Acid |
| Silver Stain Solution #6a: 4.99pH Acetate Buffer Stock | |
|---|---|
| 60 ml 1M Acetic Acid | Prepare and mix well. |
| 140 ml 1M Sodium Acetate | |
| 200 ml 4.99 pH Acetate Buffer Stock |
| Silver Stain Solution #6b: 4.99pH Acetate Buffer Working Solution | |
|---|---|
| 238 ml dH2O | Prepare and mix well. |
| 12 ml 4.99 pH Buffer | |
| 250 ml 4.99 phH Acetate Buffer Working Solution |
| Silver Stain Solution #7: 0.5% Sodium Thiosulfate | |
|---|---|
| 400 ml dH2O | Prepare and mix well. |
| 2 g Sodium Thiosulfate | |
| 400 ml 0.5% Sodium Thiosulfate |
| Silver Stain Solution #8a: Physical Developer | |
|---|---|
| Solution A | Prepare and mix well. |
| 500 ml dH2O | |
| 25 g Na2CO3 | |
| 500 ml Solution A | |
| Solution B | Prepare and mix. |
| 500 ml dH2O | |
| 1.0 g NH4NO3 | |
| 1.0 g AgNO3 | |
| 5.0 g Tungstosilicic Acid | |
| 500 ml Solution B | |
| Solution C | Prepare and mix well. |
| 250 ml dH2O | |
| 0.5 g NH4NO3 | |
| 0.5 g AgNO3 | |
| 2.5 g Tungstosilicic Acid | |
| 1.75 ml 37% Formaldehyde | |
| 250 ml Solution C | |
| Silver Stain Solution #8b: Physical Developer ABC | ||
|---|---|---|
| Free-Floating Sections | Paraffin Sections | |
| Solution A | 100 ml | 100 ml |
| Solution B | 90 ml | 80 ml |
| Solution C | 10 ml | 20 ml |