Abstract
On the observation of basophils in blood and bone marrow smears by using basic dyes, two phenomena were found and were applied in the several staining methods.
1) When fresh air-dried smears were directly dipped in the suitable high concentrated basic dye solution in methanol for the short time, the granules of basophils could be fixed and be stained concurrently (Simple staining method). Applicable dyes: Methylene blue, New methylene blue N, Toluidine blue O, Azure A and Safranin O.
2) When the suitable high concentrated basic dye solution in methanol were used for the fixation followed by the staining with aqueous basic dye solution in fresh air-dried smears, the replacement phenomen occured between the two basic dyes easily and the granules of basophils were stained securely with the latter (Replacement staining method). It was the indispensable condition that the main component of the solvent for the fixation dye is methanol and that of the staining dye is water. The dyes which could not be used in the simple staining method for the several unsuitable conditions were applied in this method. Applicable dyes: a. Fixation dyes (the dyes used in the simple staining method and Acridine orange). b. Staining dyes (Azure B, Brilliant crecyl blue, Thionin, Astra blue, Bismarck brown and etc.).
These phenomena were also common to the constituents of blood cells with the affinity to the basic dyes.
The following methods were found in the consideration for the conditions of solvents and others.
1. Astra blue (AB) staining (AB-Kernechtrot, AB-Safranin O, AB-Giemsa, AB-Periodic acid-Schiff staining).
2. Bismarck brown-Hematoxylin staining.
3. Acridine orange-Giemsa staining. (The staining mechanism of Giemsa and May-Grünwald-Giemsa staining were made clear by applying the above two phenomena.)
Using the above methods, it was considered that the clinical significances of the morphology on the basophil would be able to clear.
