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Romanowsky staining solution
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AS11L242100mL70
- Product Description
- Product FAQ
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- MSDS
- Operation video
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Product Description
Romanowsky stain consists of the basic dye azure and the acidic dye eosin, and different cellular components exhibit varying affinities for these dyes. Specifically: hemoglobin and eosinophilic granules, being basic proteins, bind to the acidic dye eosin and stain pink; nuclear proteins, lymphocytes, and the cytoplasm of basophils, being acidic, bind to the basic dyes azure or methylene blue, staining purplish‑blue or blue; neutrophilic granules, existing in an isoelectric state, can bind to both eosin and azure, resulting in a pale purplish‑red color.
Proteins are one of the major constituents of cells and exhibit amphoteric electrolytic properties, with their net charge varying according to the pH of the surrounding solution. In slightly acidic conditions, the positive charge increases, facilitating binding with eosin; erythrocytes and eosinophils stain reddish, while cell nuclei appear pale blue or remain unstained. Conversely, in slightly alkaline conditions, the negative charge predominates, promoting binding with methylene blue; all cells take on a gray‑blue hue, with granules appearing deeply dark—acidophilic granules may turn dark brown or even brown‑black—while neutrophilic granules become coarser and assume a purplish‑black color. Therefore, dilute staining solutions must be prepared using phosphate‑buffered saline (pH 6.4–6.8), and rinsing water should be near neutral; otherwise, abnormal staining may occur, making cellular morphology difficult to discern.
Order Information
Product Name
Item number
Specifications
Romanowsky staining solution
AS11L242
100mL
Transportation and Storage
Store at room temperature, protected from light. Shelf life: 12 months.
Instructions for Use
1. Smear and fix: Spread the cells evenly onto a clean microscope slide, allow them to dry, and then proceed with fixation.
The choice of fixative depends on the specific circumstances; methanol is suitable for most cells. Methanol fixation: after the smear has dried, immerse it in methanol for 15–30 minutes.
2. Dyeing:
(1) After fixation, allow the slides to air-dry at room temperature in a well-ventilated area. Place the slides in the staining tank and prepare them for staining.
(2) After adding Wright’s stain to the slide, allow it to evenly cover the cells. The exact staining time depends on the cell type; typically 2–3 minutes. It is advisable to observe the staining process under the microscope. When the Wright’s stain begins to turn reddish, promptly add an equal volume of phosphate-buffered saline (pH 6.4–6.8), and continue staining for another 2–3 minutes.
(3) After staining, first rinse the slide with phosphate-buffered saline (pH 6.4–6.8), then gently wash it with a thin stream of water to prevent large numbers of cells from being washed off the slide and compromising subsequent observations. Once rinsing is complete, place the slide in a cool, shaded area to air-dry.
3. Mounting: Mount with neutral gum arabic to prepare a permanent slide.
4. Microscopic observation: Place the dried slide under the microscope for examination.
Staining results: The cell nuclei are stained blue, while the cytoplasm is stained red.
Precautions
1. This product is intended solely for scientific research and must not be used for clinical diagnosis, treatment, or any other purposes.
2. For your safety and health, please wear a lab coat and disposable gloves when performing this procedure.
3. After the blood smear has dried, it must be fixed; otherwise, cells may easily detach during staining.
4. During rinsing, use running water; do not discard the staining solution first to prevent dye from precipitating on the blood smear. Avoid prolonged rinsing to prevent decolorization. If dye particles are deposited on the blood smear, add a few drops of methanol and immediately rinse with running water.
5. If the staining is too faint, it can be restained; when restaining, first add buffer, then add the staining solution. If the staining is too intense, rinse with running water or soak; alternatively, use methanol for destaining.
6. When using this kit for the first time, it is recommended to perform a preliminary experiment with 1–2 samples.
Related Product Recommendations
General-purpose tissue fixation solution (4% paraformaldehyde) (Catalog No.: AC28L112)
Antifade reagent (Catalog No.: AC28L512)
Wright–Giemsa Staining Solution (Catalog No.: AS11L212)
Giemsa Staining Solution (Catalog No.: AS11L232)
This product is intended solely for scientific research and must not be used for clinical diagnosis, treatment, or any other purposes.
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