Hemosiderin is a hemoglobin-derived pigment that exists as golden yellow or brownish yellow granules. It is named hemosiderin due to its iron content and golden yellow color. When red blood cells are phagocytosed by macrophages, hemoglobin is decomposed into iron-free hematoidin and iron-containing hemosiderin under the action of lysosomal enzymes. The Perls Prussian blue reaction, also known as hemosiderin staining, produces a blue color after treatment with potassium ferrocyanide and dilute acid. It is commonly found in phagocytes or interstitium and mainly demonstrates ferric iron salts. In rare cases, iron exists in its reduced ferrous form. The Lillie method is an excellent and highly specific technique for detecting ferrous iron.
L1508689
| Component | 2×50 mL | Storage | | L1508689A | Lillie Ferrous Iron Solution A
| 25 mL | RT. Store in the dark.
| | L1508689B | Lillie Ferrous Iron Solution B
| 25 mL | RT. | | L1508689C | Nuclear Fast Red Staining Solution
| 50 mL | RT. Store in the dark. |
|
Before use, mix equal volumes of Solution A and Solution B to prepare Lillie Stain. Do not prepare the stain in advance.
Operating Procedure
1. Fix tissues in 10% neutral buffered formalin or other alkaline fixatives, followed by routine dehydration and embedding.
2. Cut sections at a thickness of 4 μm, then perform routine dewaxing to water.
3. Rinse with distilled water for 1 min.
4. Immerse sections in Lillie Stain (refer to Note 4) for 25-30 min.
5. Rinse thoroughly with distilled water for 2-5 min.
6. Transfer sections to Nuclear Fast Red Staining Solution and stain nuclei lightly for 5-10 min.
7. Rinse with distilled water for 1-5 s.
8. Dehydrate and clear sections using standard protocols, then mount with neutral balsam.
Staining Results
Ferrous iron: Deep Turnbull’s blue
Cell nuclei and other tissues: Red
Precautions
1. This staining method is suitable for paraffin sections, frozen sections, and resin sections. Ensure thorough dewaxing of sections.
2. Tissues are typically fixed in 10% neutral buffered formalin. Avoid prolonged fixation in plain formalin.
3. Use clean containers throughout the procedure and avoid contact with ferrous metal products.
4. Include a single positive control section for all test sections. Selecting an appropriate control is critical for valid results.
5. Replace serial ethanol solutions regularly with fresh ones.
6. For staining frozen sections and cells, optimize experimental conditions according to specific sample types.
7. For your safety and health, wear a lab coat and disposable gloves during operation.
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