Wright's stain is a compound dye consisting of the acidic dye Eosin and the basic dye Methylene Blue, which provides good differential staining for protoplasm. Different cellular components have distinct chemical properties and varying affinities to different dyes. For instance, hemoglobin and eosinophilic granules are alkaline proteins that bind with the acidic dye Eosin and stain pink, which are referred to as eosinophilic substances. Nuclear proteins, lymphocytes and the cytoplasm of basophilic granulocytes are acidic; they combine with the basic dye to stain purplish blue or blue and are called basophilic substances. Neutrophilic granules are at the isoelectric point and can bind both Eosin and Methylene Blue, staining pale magenta and named neutrophilic substances. The cytoplasm and nucleoli of pronormoblasts and early erythroblasts contain abundant acidic substances and stain deep blue. Intermediate erythroblasts contain both acidic and alkaline substances, staining reddish blue or grayish red. Fully mature red blood cells lose all acidic substances completely and stain pink. Wright's staining solution is a clear blue liquid with a characteristic methanol odor, mainly composed of Methylene Blue, Eosin and methanol. Methanol serves to dissolve Methylene Blue and Eosin as well as fix cell morphology.
Wright's staining solution is prepared by grinding with Wright's pigment as the main raw material, which can achieve clear staining results of cells.Characteristics of this staining solution: It is composed of Wright's staining solution and phosphate buffer solution, and can be used directly without any preparation. A trace amount of neutral glycerin is added into the staining solution to prevent methanol from volatilizing or oxidizing, and also make the staining of blood cells clearer. It is often used for staining blood smears, cell smears, bone marrow cell smears and bacteria. The cytoplasm is red, cell nuclei and bacteria are blue, and eosinophilic granules are orange-red.
Component List:
| W743386 | Component | 2×100 mL | 2×500 mL | Storage | | W743386A | Wright's Stain Solution | 100 mL | 500 mL | Room temperature, store away from light | | W743386B | Phosphate Buffer Solution | 100 mL | 500 mL | Room temperatur |
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Instructions for Use:
1. Prepare blood smears, bone marrow smears or bacterial smears by conventional methods and let the smears air dry naturally.
2. Place the blood smears or bone marrow smears on a staining rack.
3. Drop an appropriate amount of Wright's staining solution to cover the smears and stain at room temperature for 2–3 minutes.
4. Add an equal volume of phosphate buffer solution onto the smears. Gently shake the slide or mix the liquids in other ways to fully blend phosphate buffer solution and Wright's staining solution, then let it stand at room temperature for 3–10 minutes.
5. Gently rinse the slide from one end with tap water or distilled water.
Note: Phosphate buffer solution can also be used to rinse the slide for about 30 seconds.
6. Air dry the slide.
7. Microscopic examination: Observe the blood smear under low-power lens first, then under oil immersion lens.
Precautions:
1. Blood smears and bone marrow smears shall be evenly spread in thickness to avoid affecting the staining effect.
2. Do not pour off the staining solution in advance or flush the slide hard during staining. Never pour out the staining solution first, otherwise dyes will precipitate and attach to the smear.
3. The staining solution can be reused but not repeatedly for many times. Filter the solution before reuse if precipitates appear.
4. Over-stained samples can be properly decolorized with methanol or ethanol; restaining is not recommended.
5. Adjust the staining time or working solution concentration if the staining result is too dark or too light.
6. pH value has a certain influence on staining. Slides must be clean and free of acid and alkali contamination to prevent poor staining results.
7. Wear a lab coat and disposable gloves during operation for your personal safety and health.
Staining Result
| Bacteria and cell nucleus | Blue | | Eosinophilic granules | Pink or orange-red | | Lymphocytes and cytoplasm of basophilic granulocytes | Purplish blue or blue | | Fully mature red blood cells | Pink |
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