Solarbio Prussian Blue Iron Stain Kit(with Neutral Red Solution)

介紹

• Hemosiderin Characteristics:
  • Derived from hemoglobin breakdown, it appears golden-yellow or brownish-yellow due to iron content.
  • Formed when macrophages engulf red blood cells and break down hemoglobin into iron-free orange blood and iron-containing hemosiderin.
• Perls Prussian Blue Reaction:
  • Also known as hemosiderin staining.
  • Produces a blue color when treated with potassium ferrocyanide and dilute acid.
  • Commonly seen in phagocytes’ interstitium, displaying ferric iron salts.
  • Classical histochemical reaction, sensitive, and excellent for displaying ferric iron in tissues.
  • The principle involves separating ferric iron from protein using potassium ferrocyanide solution and hydrochloric acid, forming an insoluble Prussian blue compound.
  • Stable ferrocyanide of ferric iron allows re-dyeing with red dye after reaction.
• Perls Stain Usage:
  • Commonly used to display hemorrhagic lesions, especially in phagocytes.
  • Helps determine hemosiderin deposition and distinguish it from other pigments.
  • The staining solution is stable, long-lasting without precipitation, and suitable for a wide range of applications.
  • Can be re-dyed, making it versatile for different staining techniques.

套件組件

使用前, mix equal parts of A1 and A-2 to form Perls Stain. It is not suitable to prepare in advance.

自我提供的材料

10% neutral formalin fixative, 系列乙醇, 蒸餾水, 4% 多聚甲醛

協定 (僅供參考)

(1) For paraffin section staining

1. Fix the tissue in 10% natural formalin fixative, dehydrate, and embed.
2. Cut sections 4µm thick, dewax to distilled water, and rinse for 1 分分鐘.
3. Soak the section in Perls Stain for 15-30 分分鐘. Rinse fully in distilled water for 2-5 分分鐘.
4. Lightly stain the nucleus with a Neutral Red Solution for 15-30 秒. Rinse in tap water for 1-5 秒.
5. Conventionally dehydrate, transparent, 並用香腸密封.

(2) For frozen section staining

1. Without dewaxing, rinse directly and quickly with distilled water for 2-3 分分鐘.
2. Follow the other steps as paraffin sections.

(3) For cultured cell staining

1. Fix in 4% 多聚甲醛用於 10-20 分分鐘.
2. Rinse in tap water twice for 2 每次幾分鐘.
3. The steps of staining, 脫水, transparency, and sealing are the same as paraffin section steps. Adjust time accordingly.

結果

• Hemosiderin or Ferric Iron: 藍色的
• Nucleus and other Tissues: Red

陰性對照

Take the same adjacent section, dewax to water. After incubation in 5% oxalic acid for 2-6 小時, follow the same procedure as above. The result should be negative.

筆記

1. Ensure clean section dewaxing.
2. 使用 10% neutral formalin for tissue fixation. Long-term fixation with common formalin can damage tissues. Avoid acid fixatives, as chromate treatment hinders iron preservation.
3. Keep containers clean and avoid using metal iron products. Use distilled water for washing sections and containers to prevent iron contamination.
4. Adjust Perls Stain dyeing time according to the sample.
5. Use the same positive control section for all sections. Autopsy lung tissue is a good control, containing many iron-positive macrophages (heart failure cells).
6. Replace ethanol series frequently.
7. For frozen section and cell staining, explore experimental conditions based on specific conditions. Wear experimental clothes and disposable gloves for health and safety.