Calcium Staining Solution (Modified Alizarin Red S Method) - BioReagent,for microscopy,Biological Stain

Cat. No.: C774863

GRADE & PURITY BioReagent, Biological Stain, for Microscopy

Synonyms

Calcium Stain Kit (Modified Alizarin Red S Method)

Storage

Store at 2-8°C,Protected from light,Room temperature

Shipped In

Wet ice

Application

Cell Staining

Overview

Calcium is abundant in the human body, forming bones that serve as the body's supporting framework, and also plays crucial roles in secretion, transport, muscle contraction, and nerve conduction. Calcium exists in the body in two forms: one is ionic calcium, present in the bloodstream (so-called blood calcium); the other is bound calcium, which combines with proteins, carbonate, or phosphate and deposits in tissues. Apart from bones and teeth, calcium normally permeates all tissues and cells and generally does not appear in a solid state within tissues. However, under certain pathological conditions, calcium can precipitate into solids and deposit within tissues, known as pathological calcification. The deposited calcium salts are primarily calcium phosphate, followed by calcium carbonate. Calcium salts are usually monorefringent, but calcium oxalate is birefringent. When stained with H&E, calcium generally appears purplish-blue. Many dyes can chelate with calcium, including Alizarin Red S, purpurin, nuclear fast red, etc. Alizarin Red S is an anthraquinone derivative, the sodium salt of alizarinsulfonic acid, which chelates with calcium salts in calcium carbonate or calcium phosphate to form an orange-red complex. Generally, these dyes are more effective in identifying moderate to large amounts of calcium than faintly stained micro-deposits, but Alizarin Red S often yields more reliable results for small amounts of deposits.

Common methods for calcium salt staining include the silver nitrate method and the Alizarin Red S method. This staining solution uses the modified McGee-Russell method with Alizarin Red S and Mayer's hematoxylin staining, particularly suitable for staining tissues with small amounts of calcium salts.

C774863	Component	3×50mL	Storage
C774863A	Alizarin Red S Staining Solution	50 mL	RT. Store in the dark.
C774863B	McGee-Russell Differentiation Solution	50 mL	RT. Store in the dark.
C774863C	Mayer's Hematoxylin Staining Solution	50 mL	2-8℃. Store in the dark.

Instructions for Use

I. Conventional Staining

1. Fix tissue in 10% neutral buffered formalin, followed by routine dehydration and embedding.

2. Cut sections at 5 µm thickness, and routinely deparaffinize to water.

3. Cover the section with Alizarin Red S Staining Solution and stain for 1~5 minutes (see Note 1). Rinse briefly with water.

4. Differentiate rapidly with McGee-Russell Differentiation Solution for a few seconds.

5. Counterstain nuclei lightly with Mayer's Hematoxylin Staining Solution for 1~2 minutes. Rinse under running tap water for 10 minutes.

6. Dehydrate and clear routinely, then mount with neutral balsam.

II. Insoluble Calcium Staining (e.g., for Undecalcified Bone)

1. Fix tissue in 10% neutral buffered formalin and dehydrate routinely. Ensure thorough dehydration in 95% ethanol.

2. Embed in plastic resin and polymerize overnight at room temperature, then in a 37°C oven for 2~4 days. Cool in a -20°C freezer for 15~20 minutes.

3. Cut sections at 3~5 µm thickness. Depolymerize undecalcified bone sections to water, then rinse with distilled water.

4. Immerse sections in Alizarin Red S Staining Solution for 1~10 minutes (see Note 2). Rinse with distilled water.

5. (Optional) Differentiate rapidly with McGee-Russell Differentiation Solution for a few seconds.

6. (Optional) Counterstain nuclei lightly with Mayer's Hematoxylin Staining Solution for 1~2 minutes. Rinse under running tap water for 10 minutes.

7. Dehydrate and clear routinely, then mount with neutral balsam.

Staining Results

Calcium Deposits	Orange-red
Cell Nuclei	Blue

Precautions

1. The staining time with Alizarin Red S should be determined based on the calcium salt content. Observe under a microscope and stop when calcium salts appear a deep orange-red, then rinse with water. Over-staining may cause diffusion; generally, 1~2 minutes is sufficient.

2. For insoluble calcium staining, the Alizarin Red S staining time should also be determined by calcium salt content. This method is commonly used in pathological diagnosis and research of pathological calcification and ossification, such as fracture repair, myositis ossificans, and calcification in old scar tissue.

3. Steps involving McGee-Russell differentiation and Mayer's hematoxylin counterstaining are not mandatory.

4. After staining with Alizarin Red S Staining Solution, calcium deposits are birefringent.

5. The Alizarin Red S method is particularly useful for identifying and detecting small amounts of calcium, such as checking for abnormal calcification in the kidney (nephrocalcinosis).

Specifications

Synonyms

Calcium Stain Kit (Modified Alizarin Red S Method)

Specifications & Purity

BioReagent, for microscopy, Biological Stain

Stability And Storage

Each component has a shelf life of 1 year under corresponding storage conditions.

Storage

Store at 2-8°C, Protected from light, Room temperature

Shipped In

Wet ice

This product requires cold chain shipping. Ground and other economy services are not available.

Grade

Biological Stain, BioReagent, for Microscopy

Chemical and Physical Properties

• Sensitivity: Light-sensitive