Giemsa stain (also known as Gimsa stain) is a mixture of Azure II and eosin. The staining principle and results of Giemsa staining are basically consistent with those of Wright's staining. Giemsa staining solution has strong staining affinity for cytoplasm, and can well demonstrate the degree of cytoplasmic basophilia. In particular, it stains the azurophilic, eosinophilic and basophilic granules in blood and bone marrow cells clearly. However, it causes over-staining of cell nuclei and poor visualization of nuclear structures. Therefore, Giemsa staining solution is often used in combination with Wright's staining solution.
Aladdin Giemsa Staining Solution is prepared by grinding, with imported Giemsa stain and methanol as the main raw materials, plus a proprietary mordant. It can produce clear cell staining results, and is commonly used for staining tissue sections, blood and cell smears, bacteria, chromosome banding, protozoan parasites, etc. Eosinophilic granules are alkaline proteins; they bind to the acidic dye eosin and stain pink, which are thus called eosinophilic substances. Nuclear proteins and lymphocyte cytoplasm are acidic; they bind to the basic dyes methylene blue or Azure and stain purple-blue, which are called basophilic substances. Neutral granules are in an isoelectric state and can bind to both eosin and methylene blue, staining pale purple, which are called neutral substances. This reagent is for research use only and is not intended for clinical diagnosis or any other purposes.
Self-prepared Materials:
Methanol, ethanol, distilled water, 0.1%–0.5% acetic acid (optional), microscope
Operating Procedures (For Reference Only):
(I) Smear Staining
1. Prepare a blood smear or bone marrow smear using standard procedures. Allow the smear to air-dry naturally, then fix it with methanol for 2–3 minutes.
2. Place the blood smear or bone marrow smear on a staining rack. Add Giemsa Staining Solution (Ready-to-use) until the smear is completely covered. Stain at room temperature or with heating for 10–15 minutes. Rinse the slide gently and slowly from one end with tap water or distilled water.
3. (Optional) Differentiate with 0.1% acetic acid for several seconds. Air-dry the smear and observe under a microscope.
Staining Results:
Eosinophilic granules
| Pink
|
Basophilic granules
| Purple-blue
|
Neutral granules
| Pale purple
|
(II) Tissue Section Staining
1. Perform routine dehydration and embedding. Cut sections at a thickness of 5 μm, then conduct routine dewaxing to water using xylene or Leagene dewaxing and clearing solution.
2. Rinse twice with distilled water, 1–2 minutes each time. Immerse the sections in Giemsa Stain for 12–24 hours, then rinse briefly with distilled water.
3. Rinse with 0.5% acetic acid for 1–2 minutes, followed by a light rinse with tap water.
4. Dehydrate rapidly three times with anhydrous ethanol, 5–10 seconds each time.
5. Clear the sections with xylene or Leagene dewaxing and clearing solution, then mount and seal with neutral balsam.
Staining Results:
Cell nucleus
| Blue to purple
|
Cytoplasmic cells
| Light blue
|
Connective tissue
| Light red
|
Precautions:
1. Blood smears or bone marrow smears shall be of uniform thickness; otherwise, the staining effect will be affected.
2. After Giemsa staining in smear staining, do not remove the staining solution first or rinse the smear vigorously directly.
3. If the staining is too dark or too light, adjust the staining time or the concentration of the staining solution accordingly.
4. In both Giemsa smear staining and tissue section staining, pH value exerts a certain influence on the staining outcome. Glass slides shall be clean and free of acid-alkali contamination; otherwise, the staining effect will be compromised.
5. If the diluted staining solution shows a metallic luster on the surface, it indicates that the solution is effective for staining; otherwise, the staining solution may have expired.
6. In Giemsa tissue section staining, after staining, rinse the sections rapidly with a large volume of 0.1%–0.5% acetic acid to avoid residual surface precipitates contaminating the sections, which are difficult to wash off subsequently.
7. Differentiation with 0.5% acetic acid is commonly used in Giemsa tissue section staining. If necessary, it can also be applied to cell smears, but its concentration should be appropriately reduced. When differentiating sections with 0.5% acetic acid, terminate the process once the sections turn pink.
8. In Giemsa tissue section staining, dehydration with anhydrous ethanol must be performed rapidly; otherwise, the sections are prone to fading.
9. The staining solution can be reused but not for excessive times. If precipitates are present, filter the solution before use.
10.For your safety and health, please wear a lab coat and disposable gloves during the operation.
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