Blood Smear Examination

Certificate in Clinical Laboratory Technology

Haematology Assignment One

Principals of Different Stains Used in Blood Smear Examination

Student ID: S150319

INTRODUCTION

Stains are used to get a clearer view of the blood cells and to find of if a patient is suffering from any type of disease. There are many types of stains which are used. However, majority of the stains come under a standard staining procedure such as Romanowsky stains. These stains have a standard staining procedure which contains acidic and a basic dye. Examples of Romanowsky stains are Giemsa stain, Leishman stain, iron stain and many more. Cytochemical stains are also one of the important which helps in identifying and confirming a myelocytic leukemia. Blood

• The stain was then mixed properly and placed the water bath of 37oc, which aided in the stain to dissolve more properly.

• Then placed the labelled bottle in the dark at room temperature.

Preparation of Leishman buffer

• The buffer was prepared by adding Sodium di-hydrogen and orthophosphate with distill water.

Staining Procedure of Leishman Stain

1. Make the blood smear and then allow it to dry.

2. Fix the smear in methanol for five minutes.

3. Flood the slide with Leishman stain and leave it for seven minutes.

4. Add Leishman buffer drop wise on the stain and leave it for twelve minutes. To allow a metallic green sheen to form on the slide.

5. Wash the slide with water and air dry.

Results of Leishman Stain

Red blood cell changed colour from red to yellowish red.

Neutrophils had pale pink cytoplasm, dark purple multi-lobed nucleus with reddish –lilac small granules.

Eosinophils were seen as blue nuclei with red to orange large granules and pale pink cytoplasm.

Basophils had dark purple to almost black large granules with purple to dark blue nucleus.

Lymphocytes had dark purple to deep bluish purple nuclei with sky blue

2) Then it is air wash for 15 min.

3) It is then stained with working solution of SBB stain solution for 1 hour.

4) With given 3 washings of ethanol for 30 sec each.

5) Washed with water and air dried.

6) Counter stained with Leishman stain.

Interpretation of the Results

• The results are similar to MPO staining both in normal and leukemic cells but the differences are:

Reference

1. Sood R. Medical Laboratory Technology: Methods and Interpretations. 6th ed. Jaypee Brothers Medical Pub; 2009.

2. Preparation of Giemsa Stain for blood film staining. http://medicine-science-and-more.com/preparation-of-giemsa-stain-for-blood-film-staining/ (accessed 19 April 2017).

3. Preparation of Leishman Stain for blood film staining. http://medicine-science-and-more.com/preparation-of-leishman-stain-for-staining-blood-film/ (accessed 19 April 2017).

4. supravitalstain.http://medicaldictionary.thefreedictionary.com/supravital+stain (accessed 20 April 2017).

5. Singh S, Veerendra Kumar S, Acharya A. Myeloperoxidase staining in the diagnosis of aggressive periodontitis. 2017.

6. [Internet]. 2017 [cited 23 April 2017]. Available from: