What should be the ideal pH for Leishman staining?
The stain must be diluted for use with Phosphate buffer to pH 6.8 or 7.2, depending on the specific technique used. 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.
What is difference between Leishman stain and Giemsa stain?
Although Giemsa staining is most commonly used, the Leishman staining method provides better visualization of the nuclear chromatin pattern of cells. It is less well known whether accuracy of parasitaemia assessment is equally accurate with the latter method.
Why Leishman stain is widely 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.
What is Leishman stain solution?
Leishman Stain Solution is used for staining blood smears to differentiate blood corpuscles, malarial parasites, trypanosomers, etc. It can be used as an alternative to Giemsa and for use in staining bone marrow sections. It serves the purpose of investigating sample material of human origin.
Why phosphate buffer is used in Leishman stain?
→ The Phosphate buffer has an extremely high buffering capacity and gives better results in Hematology staining.
Why buffer is used in Leishman stain?
In that case the buffer solution is essential for preparation of diluted Giemsa/May-Gruenwald/Wright/Leishman solutions and for rinsing stained samples without causing destaining of stained cells. Buffer solutions are solutions of weak acids and theirs salts or weak bases and their salts.
What is solution for Leishman stain?
It is based on a methanolic mixture of “polychromed” methylene blue. (i.e. demethylated into various azures) and eosin. The methanolic stock solution is stable and also serves the purpose of directly fixing the smear eliminating a prefixing step. Quality Control.
What are the reagents of Leishman powder?
Leishman stain is a mixture of Methylene blue (Basic), and Eosin (Acidic) dye.
How do you make a phosphate buffer for Leishman stain?
⇒ Measure the 800 ml of distilled water with the help of measuring cylinder & Pour it into a beaker. ⇒ To this, add 20.209 grams of Sodium phosphate dibasic and mix well with the help of stirrer. ⇒ Now add 3.394 grams of Sodium phosphate monobasic and mix well using the glass stirrer.
What is the fixative present in Leishman stain?
The fixative present in Leishman stain is Buffered formalin J10% formalin ~Ethy alcohol ~Acetone free methanol 02.
What happens if the buffer used in Leishman’s staining is highly acidic?
Excessive acidic pH or reduced staining time causes excessive pinkish discoloration of smear, while excessive basic pH and/or increased staining time causes bluish discoloration.