Solarbio Prussian Blue Iron 얼룩 키트(뉴트럴 레드 솔루션으로)

소개

• 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.

키트 구성품

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

자체 제공 자료

10% neutral formalin fixative, 일련의 에탄올, 증류수, 4% 파라 포름 알데히드

규약 (참고용으로만)

(1) For paraffin section staining

1. 티슈를 고정하세요. 10% natural formalin fixative, dehydrate, and embed.
2. Cut sections 4µm thick, dewax to distilled water, and rinse for 1 분.
3. Soak the section in Perls Stain for 15-30 분. Rinse fully in distilled water for 2-5 분.
4. Lightly stain the nucleus with a Neutral Red Solution for 15-30 초. Rinse in tap water for 1-5 초.
5. Conventionally dehydrate, transparent, 그리고 레진으로 밀봉하세요.

(2) For frozen section staining

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

(3) For cultured cell staining

1. Fix in 4% 파라 포름 알데히드 10-20 분.
2. Rinse in tap water twice for 2 매번 분.
3. The steps of staining, 탈수, transparency, and sealing are the same as paraffin section steps. Adjust time accordingly.

결과

• Hemosiderin or Ferric Iron: 파란색
• Nucleus and other Tissues: 빨간색

부정적인 제어

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

메모

1. Ensure clean section dewaxing.
2. 사용 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.