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

การแนะนำ

• 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.

ส่วนประกอบชุดอุปกรณ์

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

วัสดุที่จัดหาด้วยตนเอง

10% neutral formalin fixative, Series of ethanol, น้ำกลั่น, 4% paraformaldehyde

มาตรการ (สำหรับการอ้างอิงเท่านั้น)

(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 นาที.
3. Soak the section in Perls Stain for 15-30 นาที. Rinse fully in distilled water for 2-5 นาที.
4. Lightly stain the nucleus with a Neutral Red Solution for 15-30 วินาที. Rinse in tap water for 1-5 วินาที.
5. Conventionally dehydrate, transparent, and seal with resinene.

(2) For frozen section staining

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

(3) For cultured cell staining

1. Fix in 4% พาราฟอร์มัลดีไฮด์สำหรับ 10-20 นาที.
2. Rinse in tap water twice for 2 นาทีในแต่ละครั้ง.
3. The steps of staining, dehydration, transparency, and sealing are the same as paraffin section steps. Adjust time accordingly.

ผลลัพธ์

• Hemosiderin or Ferric Iron: สีฟ้า
• Nucleus and other Tissues: สีแดง

การควบคุมเชิงลบ

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

บันทึก

1. Ensure clean section dewaxing.
2. ใช้ 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.