The Standard Operating Procedures (SOPs) are different for almost every laboratory as per the quality of the staining solution & buffer they are using and the purpose of staining. The pH 6.8 is preferred when the morphology of blood cells is to be examined and pH 7.2 is good for parasitic studies. The stain must be diluted for use with Phosphate buffer to pH 6.8 or 7.2, depending on the specific technique used. and the Basic dye, Methylene blue stains the Acidic components, especially the Nucleus of the cell. The acidic dye, Eosin, variably stains the Basic components of the cells i.e. As it is a type of Romanowsky stains, it contains both the Acidic and Basic dyes which have the affinity for Basic and Acidic components of the Blood cells respectively. The working principle of the Leishman stain is same as described above. This stain is the most dependable stain for Peripheral blood film examination. and also used to differentiate nuclear and cytoplasmic morphology of the various cells of the blood like Platelets, RBCs, WBCs as well as for the parasites. Leishman stain is commonly used when there is need to examine the Blood smear for the Various blood cells, Differential Leucocyte count, Type of Anemia, Toxic Granules & Platelet count etc. The methanolic stock solution is stable and also serves the purpose of directly fixing the smear eliminating a prefixing step. It is based on a methanolic mixture of “polychromed” Methylene blue and eosin. It stains the different components of blood in a range of shades between red and blue. It consists of a mixture of eosin (an acidic stain), and Methylene blue (a basic stain) in Methyl alcohol and is usually diluted and buffered during the staining procedure. Leishman Stain is a neutral stain for blood smears which was devised by the British surgeon W. Giemsa stain gives better results in parasitic studies. The Leishman stain is one of the best stains for routine blood stain to stain the Peripheral blood smear for the examinations of blood film under the microscope and is satisfactory for malaria and other blood parasites. It differentially stains the human and bacterial cells and appeared as purple and pink colored bodies respectively. Leishman stain is a differential stain that is used to variably stain the various components of the cells and it can be used to study the adherence of pathogenic bacteria to the human cells. Leishman stain is a mixture of Methylene blue, and Eosin dye, prepared in Alcohol medium and diluted with buffer or distilled water during staining procedure. In this Article, I’ll discuss Leishman Staining Technique used in Hematology Laboratory. There are 4 different types of Romanowsky stains commonly used in Hematology laboratory for staining the blood cells – Most of the Romanowsky stains are prepared with Methyl alcohol (Methanol) so that they act as a fixative as well as the cellular stain.
the cytoplasm and Granules in some cells. Nucleus and Eosin/ Azure is the Acidic dye which has the high affinity for the basic components of the cells i.e. Methylene blue is a basic dye that has a high affinity for the acidic components of the cell i.e. Almost all the Romanowsky group of stains has two essential components i.e. Romanowsky stains are such types of stains that are universally employed for the staining of blood cells.
Besides the dyes, a buffer is added to the stain which acts as the mordant and enhances the staining reaction, results in the better morphology of the blood cells under the microscope. Therefore, the stains used in Hematology laboratory are the combination of these three types of stains. they have a high affinity for Basic Dyes and then there are some components of the cells which are neutral and has the high affinity for neutral stains. they have a great affinity for acidic dyes whereas some of the components are Acidophilic i.e. Some of the Components of the blood cells are Basophilic i.e. The color of Granules by Leishman StainĪs you already know, that there are various types of Blood cellspresent in our body, having different shapes and sizes, which take up the stains as per their structures.The color of Cytoplasm by Leishman Stain.Commonly the Covering Technique is employed for staining the Thin blood smears with Leishman stain & here I’m explaining the Leishman staining technique for thin blood smear by Covering method….MATERIALS REQUIRED FOR THE LEISHMAN STAIN.A brief Description of Leishman Stain….
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