Difference Between Giemsa Stain and Leishman 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 and identify trypanosomes, leucocytes and malaria parasites.

Staining is a vital step during the enhancement of the contrast of a microscopic image in the context of microscopy, especially to highlight different structures in biological cells and tissues. Giemsa stain and Leishman stain belong to the group of Romanowsky stains, which also include Wright stain and Jenner stain. Commonly, Romanowsky stains are useful in staining blood smears. We mainly use this during the study of red blood cell morphology and the performance of differential white blood cell counts. Eosin Y and azure B dyes are the common components for Romanowsky stains. Romanowsky staining procedures help in the diagnosis of different disease conditions such as Leukemia.

CONTENTS

1. Overview and Key Difference
2. What is Giemsa Stain
3. What is Leishman Stain
4. Similarities Between Giemsa Stain and Leishman Stain
5. Side by Side Comparison – Giemsa Stain vs Leishman Stain in Tabular Form
6. Summary

What is Giemsa Stain?

Giemsa stain usually helps to differentiate between cytoplasmic and nuclear morphology of blood cells like white blood cells, red blood cells and platelets. Also, it helps to differentiate the parasites. Mainly, it is used for cytogenetics and in the diagnosis of malaria and other parasitic diseases. Moreover, Giemsa stain is specific for the phosphate groups which are present in the DNA. They adhere to the areas where there is a higher number of adenine-thymine bonding in the DNA strand.

Furthermore, Giemsa stain is also useful in Giemsa banding or G banding to stain chromosomes and to generate karyograms. Therefore, Giemsa stain has the ability to identify and visualize different abnormalities in the chromosomes. For example, the trophozoite of Trichomonas vaginalis, which gives out a greenish discharge and consists of motile cells on wet preparations, is stained with Giemsa stain. As mentioned above, Giemsa stain acts as a typical blood film stain. When stained, red blood cells stain in pink, platelets stain in light or pale pink, and the cytoplasm of lymphocytes, monocytes and leukocytes stain in sky blue, pale blue and magenta, respectively.

Giemsa stain is a mixture of eosin, methylene blue and Azure B. The methylene azure mixture which forms an eosinate along with methylene blue stabilizes this mixture. In the process of Giemsa stain, a thin film of the specimen is placed on a microscopic slide first. The next step is fixing with pure methanol for about 30 seconds by adding a few drops of methanol on the slide. Then, the slide is immersed in the 5% Giemsa stain solution for about 20-30 minutes. The final step is washing the slide with tap water and leaving to dry.

What is Leishman Stain?

The Scottish pathologist William Boog Leishman is the developer of Leishman stain. It is one of the stains that belongs to the group of Romanowsky stains. Moreover, it is utilized more commonly in the differentiation and identification of different malarial parasites, trypanosomes – a unicellular flagellated protozoa, which are parasitic and leucocytes.

The basis of Leishman stain is a methanolic mixture that contains a mixture of methylene blue which is ‘polychromed’; demethylated into different types of azures and eosin. Due to the stability of the stock solution of the methanolic mixture, we can use it directly into fixing the smear, while eliminating the prefixing step. The stability decreases if the solution is mixed with an aqueous buffer. When performing differential cell counts, Leishman stain provides characteristic bright violet colour to the nucleus and neutrophil granules. Thus, it enhances the differentiation between the nucleus and the cytoplasm. Also, the Leishman stain provides more quality contrastive staining in comparison with other stains that are methylene blue and eosin based.

Since different cytoplasmic components are addressed sharply for differentiation and identification, haematologists prefer Leishman stain in comparison to other Romanowsky stains. In the detection of malarial parasites, Leishman staining procedures are more sensitive and accurate than other stains such as Field’s stain.

What are the Similarities Between Giemsa Stain and Leishman Stain?

• Giemsa stain and Leishman stain are differential stains.
• Both are useful during the performing of differential white blood cell counts and study of cell morphology of red blood cells.
• Moreover, both stains belong to the group of Romanowsky stains.

What is the Difference Between Giemsa Stain and Leishman Stain?

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 in blood staining to differentiate and identify trypanosomes, leucocytes and malaria parasites. So, this is the key difference between Giemsa stain and Leishman stain.

Moreover, bacteriologist Gustav Giemsa developed the Giemsa staining technique, while pathologist William Boog Leishman developed Leishman staining technique. Further, another significant difference between Giemsa stain and Leishman stain is the composition of the stain. Giemsa stain is a mixture of eosin, methylene blue and Azure B while Leishman stain is a methanolic mixture that contains a mixture of methylene blue.

Summary – Giemsa Stain vs Leishman Stain

In the context of microscopy, staining techniques play a major role in enhancing the contrast of microscopic images of various biological tissues. Giemsa staining is useful in the staining of DNA regions of different chromosomes to investigate different aberrations such as translocations and rearrangements. Leishman stain is useful in blood smear staining and analysis to differentiate and identify trypanosomes, leucocytes, and malaria parasites. So, this is the difference between Giemsa stain and Leishman stain.

 Reference:

1. Gajendra, S, et al. “Leishman and Giemsa stain: a new reliable staining technique for blood/Bone marrow smears.” International journal of laboratory hematology., U.S. National Library of Medicine, Dec. 2015, Available here. Accessed 13 Sept. 2017.
2. Sathpathi, Sanghamitra, et al. “Comparing Leishman and Giemsa staining for the assessment of peripheral blood smear preparations in a malaria-Endemic region in India.” Malaria Journal, BioMed Central, 2014, Available here. Accessed 13 Sept. 2017.

Image Courtesy:

1. “Trypanosoma cruzi crithidia” By CDC/Dr. Myron G. Schultz –  from the Centers for Disease Control and Prevention’s Public Health Image Library (PHIL) (Public Domain) via Commons Wikimedia
2. “Plasmodium vivax malaria” By Department of Pathology, Calicut Medical College – Calicut Medical College (CC BY-SA 4.0) via Commons Wikimedia