What is the preparation of Leishman stain?

What is the preparation of Leishman stain?

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.

How do you make a buffer solution for Leishman stain?

Methanol in the stain fixes the preparation. 3) Add double the volume of distilled water to the slide and mix. 4) Allow the diluted stain to act for 10-12 minutes. 5) Wash the film with distilled water or phosphate buffer of pH 7.0, drain and dry in air and examine.

How blood smear is prepared?


  1. A small drop of blood is placed on the midline at the end of a glass slide.
  2. Second slide (ideally narrower than smear slide, to avoid spreading the cells over the edge) is placed on the smear slide in front of the blood spot in a way that it forms a 30-45°angle on the side of the blood drop.

Why is Leishman stain used?

Leishman stain, also known as Leishman’s stain, is used in microscopy for staining blood smears. It is generally used to differentiate between and identify white blood cells, malaria parasites, and trypanosomas.

Which stain is used in DLC?

Generally, methylene blue or touline are basic stains, and Eosin, Azure-I, Azure-II are the acidic stains in use. It is advisable to stain a slide soon after preparation of blood smear.

Why is Leishman Stain used?

Leishman stain is used in microscopy for staining blood smears. It provides excellent stain quality. It is generally used to differentiate and identify leucocytes, malaria parasites, and trypanosoma. It is based on a methanolic mixture of “polychromed” methylene blue.

Is Leishman Stain a romanowsky stain?

Stains that are related to or derived from the Romanowsky-type stains include Giemsa, Jenner, Wright, Field, May–Grünwald and Leishman stains.

Why is Leishman stain diluted after two minutes and not earlier?

This will stain the blood cells. ⇒ Rinse the slides thoroughly with Phosphate buffer solution up to 2 minutes or until it acquires a purple-pinkish tinge. ⇒ Air dry the slides in a tilted position so that the water easily remove out of the slides. The color of Cytoplasm by Leishman Stain.

What are the two types of smear preparation?

Four different types of smear preparation methods (conventional method, blood film method, drop and rest method, and water-wash method) were carried out according to the standard reference as described below. A drop of sample was placed onto a clean slide and then spread to make a smear of 1 cm diameter.

Why is thick smear not fixed?

Thick smear. It is not fixed in methanol; this allows the red blood cells to be hemolyzed, and leukocytes and any malaria parasites present will be the only detectable elements. However, the hemolysis may lead to distorted plasmodial morphology making plasmodium species differentiation difficult.

What is the difference between Leishman Stain and Giemsa stain?

The key difference between Giemsa Stain and Leishman Stain is that Giemsa staining is useful in the staining of DNA regions of different chromosomes to investigate different aberrations such as translocations and rearrangements, while Leishman stain is useful during blood smear staining and analysis to differentiate …

Can a Leishman stain be prepared in a laboratory?

Nowadays, commercially prepared Leishman staining solutions are used in most of the laboratories. However, some laboratories still prepare it in their own setup as per their Standard Operating Procedure (SOP). Here, I’m gonna explain the method of Preparing Leishman Stain or Leishman Stock solution in Laboratory.

Which is better Leishman stain or Giemsa stain?

Giemsa stain gives better results in parasitic studies. Leishman Stain is a neutral stain for blood smears which was devised by the British surgeon W. B. Leishman (1865–1926). 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.

How is methanolic stock used to stain blood?

It stains the different components of blood in a range of shades between red and blue. It is based on a methanolic mixture of “polychromed” Methylene blue and eosin. The methanolic stock solution is stable and also serves the purpose of directly fixing the smear eliminating a prefixing step.

What kind of alcohol is used to prepare Romanowsky stain?

Most of the Romanowsky stains are prepared with Methyl alcohol (Methanol) so that they act as a fixative as well as the cellular stain. There are 4 different types of Romanowsky stains commonly used in Hematology laboratory for staining the blood cells –