Solarbio Von Kossa Calcium Stain Kit

Kit Components

Introduction

Calcium is abundant in the human body, forming bones as a scaffold to support it. Calcium plays a crucial role in secretion, transportation, muscle contraction, nerve conduction, and more. Several dyes can form chelates with calcium, such as Alizarin Red 5, Purpurin, Nuclear Fast Red, etc. Alizarin Red S, a sodium alizarin sulfonate, is an anthraquinone derivative capable of chelating with calcium carbonate or calcium phosphate to form an orange-red complex. Common methods for calcium salt staining include the Von Kossa method and the Alizarin Red S method. This kit’s principle relies on a metal replacement method: when Von Kossa Silver Solution acts on sections containing insoluble calcium salt, calcium is replaced by silver, and the silver salt reduces to black metal silver under light exposure. It’s suitable for calcium salt tissue staining of numerous samples.

Protocol (for reference only)

1. Fix the tissue with 10% NBF and conventionally dehydrate and embed.
2. Cut the section into 5µm, conventionally dewax and hydrate.
3. Dye the section with Von Kossa Silver Solution under strong light for 15–60 minutes.
4. Rinse with distilled water for 1 minute. Treat with Hypo Solution for 2 minutes.
5. (Optional) Re-dyeing with HE solution or Van Gieson Solution.
6. Conventionally dehydrate, transparent, and seal with resinene.

Negative Control (optional)

Take another adjacent section and dewax it to water. Put it into Von Kossa Control Solution for 20 minutes, then rinse it with running water for 5 minutes. Subsequently, place it into Von Kossa Silver Solution together with the experimental section, and follow the same steps as above.

Result

• Calcium Salt: Brownish Red to Dark Black
• Nucleus: Range with re-dyeing solution
• Background: Range with re-dyeing solution

Note

1. It’s best to use NBF for fixing tissues containing calcium salt.
2. The action time depends on the intensity and duration of light exposure.
3. If exposed to UV light, reduce exposure time to 5-10 minutes.
4. For safety and health, wear experimental clothing and disposable gloves