Prussian blue staining solution (nuclear fixation method)
Product introduction:
Hemosiderin (Hemosiderin) is a hemoglobin-derived pigment, which is a golden yellow or brownish yellow granule. Because it contains iron and is golden yellow, it is called hemosiderin. When red blood cells are phagocytosed by macrophages, hemoglobin is decomposed into iron-free orange blood and iron-containing hemosiderin under the action of lysosomal enzymes. Perls Prussian blue reaction, also known as hemosin staining, can produce blue after treatment with potassium ferrocyanide and dilute acid, which is common in phagocytic cells or interstitial cells. The potassium cyanide solution separates ferric ions from the protein by dilute hydrochloric acid, and the ferric iron reacts with potassium ferrocyanide to form an insoluble blue compound, ferrocyanide of ferric iron.
NobleRyder Prussian blue staining is used to show various hemorrhagic lesions in local tissues, which are common in phagocytic cells and can distinguish well from hemosiderin and other pigments. The dyeing solution has good stability, can be stored for a long time, is not easy to precipitate, has a wide application range, and can be counterstained. The counterstaining solution of the dyeing solution adopts nuclear solid red, which is the most classic and most commonly used counterstaining solution.
Product composition :
Numbering name | D3801 2 x 50ml | D3801 2×100ml | Storage | |||
Reagent (A): | A1: Perls stain A | 25ml | 50ml | RT | ||
Perls stain | A2: Perls stain B | 25ml | 50ml | RT | ||
Before use, take the same amount of A1 and A2. | Perls stain, should not be prepared in advance. | |||||
Reagent (B): Nuclear solid red staining solution | 50ml | 100ml | RT protected from light | |||
user's Guide | 1 copy |
Bring your own materials:
1. Fixative solution: 10% neutral formalin, 4% paraformaldehyde, etc.
2, series of ethanol
Operation steps (for reference only):
(1) Paraffin section staining:
1. The tissue is fixed in 10% neutral formalin and conventionally dehydrated and embedded.
2. Slice thickness 4 μm, conventional dewaxing to water.
3. Wash the distilled water for 1 min.
4. Slice into Perls stain (see Note 2) and dip for 15 to 30 minutes.
5. Rinse the distilled water for 2 to 5 minutes.
6, into the nuclear red staining solution, lightly stained nuclei 5 ~ 10min.
7. Rinse tap water for 1 to 5 seconds.
8, conventional dehydration transparent, neutral gum sealing
(2) Frozen section dyeing:
1. Without dewaxing, rinse directly with distilled water for 2 to 3 minutes.
2, dyeing, dewaxing, transparent, sealing steps with the staining steps of paraffin sections, the time can be shortened accordingly.
(3) Cell staining:
1, 4% paraformaldehyde fixed for 10 ~ 20min.
2, tap water 2 times, each time 2min.
3. Rinse the distilled water twice for 2 minutes each time.
4, dyeing, dewaxing, transparent, sealing steps with the staining steps of paraffin sections, the time can be shortened accordingly
Dyeing results:
Hemosiderin or ferric iron | blue |
Nucleus, other tissues | red |
Negative control (optional): The same section was dewaxed to water. Incubate with 5% oxalic acid for 2-6 hours, pass Perls stain, and the rest of the steps are the same as above. The result was negative.
Precautions:
1, slice dewaxing should be as clean as possible. Tissue fixation often uses 10% neutral formalin, which is damaged by long-term fixation with common formalin. Avoid the use of acidic fixatives, and chromate treatment can also hinder the preservation of iron.
2. The container should be clean during the whole operation process, avoiding the use of metal iron products, and it is better to use distilled water when washing slices and containers, because ordinary water contains iron. When staining Perls stain, the coloring time should be adjusted according to the sample condition.
3. The same positive control section should be used for all sections. It is important to select a suitable control. Autopsy lung tissue is a good control and contains a significant number of iron-positive macrophages (heart failure cells).
4, frozen section and cell staining, it is best to explore the experimental conditions according to the specific situation.
Validity: Valid for 12 months.
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